PEPTIDE HORMONE WITH ONE OR MORE O-GLYCANS
20210171598 · 2021-06-10
Assignee
Inventors
- Katrine TER-BORCH GRAM SCHJOLDAGER (Vedbaek, DK)
- Henrik CLAUSEN (Copenhagen Ø, DK)
- Sergey Y. VAKRUSHEV (Nyborg, DK)
- Christoffer Knak GOTH (Frederiksberg C, DK)
- Thomas Daugbjerg MADSEN (Copenhagen N., DK)
- Lasse H. HANSEN (Glostrup, DK)
- Jens P. GØTZE (Virum, DK)
Cpc classification
C07K1/006
CHEMISTRY; METALLURGY
C07K19/00
CHEMISTRY; METALLURGY
A61K38/29
HUMAN NECESSITIES
C07K1/1072
CHEMISTRY; METALLURGY
International classification
C07K1/00
CHEMISTRY; METALLURGY
Abstract
The present invention relates to a peptide hormone with one or more O-glycans attached at specific amino acid residues as well as to formulations comprising the same.
Claims
1. A formulation comprising at least one molecule of a peptide hormone species exhibiting a specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone species, wherein specific, defined glycosylation pattern means that the each molecule of said peptide hormone in said pharmaceutical formulation displays structural homogeneity with respect to the site of glycan attachment and/or with respect to the glycan attachment.
2. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 1, 2, 3, 5, 6, 7, 14, 15, 16, 21, 22, 23, 24, 25, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49, 51, 52, 54, 55, 56, 57, 72, 73, 74, 76, 79, 8, 81, 83, 84, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 106, 107, 108, 109, 113, 116, 117, 118, 119, 120, 130, 131, 135, 136, 143, 144, 147, 163, 164, 167, 168, 170, 184, 185, 186, 188, 189, 190, 191, 192, 204, 215, 217, 219, 222, 227, 229, 230, 231, 233, 234, 236, 252, 260, 262, 267, 272, and/or 279.
3. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 6, 7, 21, 22, 23, 24, 72, 74, 92, 95, 97, 99, 106, 107, 108, 116, 117, 118, 147, 163, 167, 185, 186, and/or 188.
4. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 72, 95, 97, 106, 108, 147, 185, and/or 188.
5. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone species is SEQ ID NO: 147.
6. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said predetermined specific site of said peptide hormone is within the receptor-binding region.
7. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone exhibiting an specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone is obtainable by recombinant production using a host cell in which one or more glycosyltransferase gene(s) is/are inactivated or downregulated by inactivation and/or downregulation of one or more gene(s) selected from COSMC and C1GALT1; and/or by inactivation and/or downregulation of one or more gene(s) selected from GCNT3, GCNT4, B3GNT6, and/or by inactivation and/or downregulation and/or upregulation/activation of one or more gene(s) selected from ST6GALNAC1-6, ST3GAL1, GCNT3, GCNT4, and/or B3GNT6.
8. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein said peptide hormone species exhibits a specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone and, wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with optional elongation and sialic acid capping, wherein optionally the first monosaccharide attached in the synthesis of O-linked glycans is N-acetyl-galactosamine and wherein a core1 structure may be obtained by the addition of galactose, and wherein a core2 structure may be obtained by the addition of N-acetyl-glucosamine to the N-acetyl-galactosamine of the core1 structure and wherein the core3 structures may be obtained by the addition of a single N-acetyl-glucosamine to the first monosaccharide N-acetyl-galactosamin and core4 structures may be obtained by the addition of a second N-acetyl-glucosamine to the core3 structure.
9. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
10. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
11. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
12. A formulation comprising at least one molecule of a peptide hormone species according to claim 1, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
13. A formulation comprising at least one molecule of a peptide hormone species according claim 1, wherein the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
14. A formulation according to claim 1, wherein said formulation is a pharmaceutical formulation.
15. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 1, wherein said peptide hormone species comprises one or more O-linked glycan at a site indicated in and one of Tables 6A to 6E, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
16. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site indicated in Table 6A, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
17. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6B, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
18. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
19. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
20. A formulation comprising at least one molecule of a peptide hormone species as defined in claim 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6D, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
21. A modified peptide hormone comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6A, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 1, 2, 3, 5, 6, 7, 14, 15, 16, 21, 22, 23, 24, 25, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49, 51, 52, 54, 55, 56, 57, 72, 73, 74, 76, 79, 8, 81, 83, 84, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 106, 107, 108, 109, 113, 116, 117, 118, 119, 120, 130, 131, 135, 136, 143, 144, 147, 163, 164, 167, 168, 170, 184, 185, 186, 188, 189, 190, 191, 192, 204, 215, 217, 219, 222, 227, 229, 230, 231, 233, 234, 236, 252, 260, 262, 267, 272, and/or 279.
22. The modified peptide hormone comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6B, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 6, 7, 21, 22, 23, 24, 72, 74, 92, 95, 97, 99, 106, 107, 108, 116, 117, 118, 147, 163, 167, 185, 186, and/or 188.
23. The modified peptide hormone comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 72, 95, 97, 106, 108, 147, 185, and/or 188.
24. The modified peptide hormone comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6D, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is depicted in SEQ ID NO: 147.
25. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with sialic acid capping, wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with optional elongation and sialic acid capping, wherein optionally the first monosaccharide attached in the synthesis of O-linked glycans is N-acetyl-galactosamine and wherein a core1 structure may be obtained by the addition of galactose, and wherein a core2 structure may be obtained by the addition of N-acetyl-glucosamine to the N-acetyl-galactosamine of the core1 structure and wherein the core3 structures may be obtained by the addition of a single N-acetyl-glucosamine to the first monosaccharide N-acetyl-galactosamin and core4 structures may be obtained by the addition of a second N-acetyl-glucosamine to the core3 structure.
26. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
27. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
28. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
29. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
30. The modified peptide hormone according to claim 21, wherein the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
Description
LEGENDS TO THE FIGURES
[0035]
[0036]
[0037]
[0038]
[0039]
[0040]
[0041]
[0042]
[0043]
DETAILED DISCLOSURE OF THE INVENTION
[0044] O-glycosylation is emerging as an important regulator of protein stability and function. In this study, for the first time, the present inventors identify protein O-glycosylation as a common post translational modification of peptide hormones, present a detailed comprehensive map of O-glycosylation sites and suggest a biological function of site-specific O-linked glycosylation on peptide hormones.
[0045] Peptide hormones are produced by cells of the endocrine, neuronal or neuroendocrine tissues and are secreted in response to stimulus to bind to their cognate receptors and regulate complex physiological processes like appetite, blood pressure and anxiety. During biosynthesis peptide hormones undergo a range of PTMs. Besides a common complex proprotein convertase activation.sup.19, peptide hormones can undergo C-terminal amidation, N-terminal acetylation, tyrosine sulfation and serine phosphorylation.sup.20 that may change the biochemical properties of the peptides. Furthermore, peptide hormones circulate in minute amounts and are inherently prone to proteolytic degradation. Thus, as a result of their instable nature, low abundance and complex post translational modifications peptide hormones have been difficult to isolate and characterize.
[0046] Now with the advantage of sensitive mass spectrometry the present inventors explore the occurrence of O-glycans on peptide hormones and show that approximately one third of all classified (NeuroPeP) peptide hormones are O-glycosylated (
[0047] GalNAc O-glycosylation is an exceptional PTM in that there are 20 differentially expressed isoforms with partly overlapping specificities conducting the addition of the initial GalNAc residue to the protein backbone Ser/Thr/Tyr residues. This leaves ample room for regulating the addition of site-specific O-glycosylation and the findings presented here may have revealed a novel regulatory level in peptide hormone biosynthesis and function.
[0048] It is becoming increasingly clear that site-specific O-glycosylation fine-tune the biological function of proteins.sup.21-32. Previously the present inventors have demonstrated that O-glycosylation protects proproteins from PC processing.sup.24,33,34, that O-glycans increase the stability of GPCR N-termini.sup.32 and modulates ectodomain shedding.sup.26 and most recently the present inventors have shown that loss of site-specific O-glycosylation impair ligand binding and uptake of the LDLR related receptor family.sup.35,36.
[0049] Here the present inventors demonstrate that O-glycosylation of VIP, Secretin, NPY, PYY and PPY in specific well conserved amino acid positions lowers receptor affinity and signalling, reducing EC50 more than 28-fold where the size of the glycan correlates with signal reduction (
[0050] Our data indicates that the presence of glycan structures somehow alters the interaction between peptide hormone ligand and cognate receptor. Well in line with these data, especially well-studied in the Neuropeptide Y family, it has been demonstrated that other bulky chemical modifications of amino acids in the receptor binding domain alter receptor sub-class selectivity of the ligand.sup.37. Such a sub-class receptor selectivity was observed for Thr32-Tn modified NPY that retained activity at the NPY2R and NPY5R receptors with a 118-fold and 37-fold decrease in potency (
[0051] Peptide hormones are inherently unstable and circulate only in minute amounts. Here, it is demonstrated that site-specific O-glycosylation on Secretin, VIP, Galanin, PYY, GLP-1 and ANP protects the peptide hormones from proteolytic degradation in vivo using a rat model, ex vivo using plasma degradation assays and in vitro using recombinant proteases (
[0052] The Neuropeptide Y family members are ubiquitously expressed in the body and act as neurotransmitters to regulate a vast array of physiological processes via binding and signalling through the Gi coupled NPY receptors (YR1, YR2, YR4 or YR5).
[0053] The main function of GLP-1 is to increase insulin secretion, i.e. to act as an “incretin”, but it also inhibits gastrointestinal motility and function as a physiological regulator of appetite. Recently it was demonstrated that GLP-1, Oxyntomodullin and PYY in combination injected subcutaneously using a pump device into obese volunteers reduced their mean caloric intake with 32%.sup.38.
[0054] ANP is classically released from secretory granules from the atria in a regulated fashion which makes it able to rapidly regulate hemodynamics in response to increased pressure. Glycosylated ANP was protected from degradation in vitro by IDE and NEP which suggest that glycosylated ANP may have increased half-life. The combined effects of glycosylated ANP, i.e decreased potency and increased stability, could render a positive effect in the treatment of acute decompensated heart failure and hypertension, where ANP and BNP are already introduced as infusion therapy.sup.39-41.
[0055] Thus, there is a need to identify selective peptide hormone receptor agonists or antagonists with good biostability to pursue as potential therapeutic candidates for treating e.g. cardiovascular diseases, anxiety, depression, obesity, epilepsy, alcoholism.
[0056] Peptide based design of therapeutics is attractive in many ways since biological active peptides have the potential to regulate specific functions of GPCRs and ion-channels and in general in vivo based drug design is favourable due to lower toxicity and immunogenicity and higher selectivity and predictable in vivo behaviour. However, probing the function and efficacy of what was thought of as “naturally occurring” unmodified peptides have demonstrated low biostability and circulation time and therefore low efficacy.sup.42,43 and a number of strategies have been taken to chemically alter the biochemical properties of peptide hormones or synthetic analogues and improve these parameters.sup.44. It is generally found that large N-glycans on proteins may enhance circulatory half-life, although mainly by increase of the size and hydrodynamic size of proteins. However, introduction of O-glycans into smaller peptides have also been used to enhance circulatory half-life especially when combined with the GlycoPegylation strategy.sup.45. In one relevant example an O-glycosylation sequon was introduced in the C-terminus of the GLP-1 receptor antagonist Exendin (9-39) increasing functional half-life.sup.46. Other studies have explored the chemical synthesis of peptide hormones with glycans.sup.46-52, e.g. O-linked galactose on vasopressin, PYY and VIP, glucose on Leu-enkephalin and PYY, N-linked GlcNAc on GLP-1 and N-terminal chemical glycation of GIP, GLP-1(7-36) and Insulin. Interestingly all studies, except one describing O-linked galactose on Vasopressin, find, very much in line with what the present inventors show here for the naturally occurring O-GalNAc glycans, that the chemically modified glycosylated analogues are less or equally potent in in vitro receptor activation assays, yet these other studies demonstrate higher stability and potency in vivo.
[0057] Naturally occurring O-glycans positioned within the receptor-interaction region of peptide hormones have not been demonstrated previously, and we hypothesize that such naturally occurring glycans selectively affect function and biostability as our preliminary studies suggest.
[0058] The inventors of the present invention also identified O-glycosylation on the pro-part of peptide hormones. Here, sequestered from the bioactive part, the sugars might regulate PC processing and activation and furthermore coincidentally mask antibody binding epitopes.sup.9,24,53-56. One such well-studied example is proBNP which is synthesized in the ventricles of the failing heart and undergo limited proteolysis by PCs releasing the C-terminal peptide hormone BNP that regulate natriuresis and blood pressure. ProBNP is O-glycosylated in the N-terminal proprotein (NT-proBNP) close to the PC processing site and amino acid substitution experiments have validated that Thr71 protects proBNP from processing by Corin or Furin.sup.53. The present inventors identified O-glycans on proBNP, POMC, Kininogen-1, Chromogranin A (Table 6). NT-proBNP is an important biomarker for heart failure and commercial immunoassays are being used in the clinic to quantify NT-proBNP in heart related diseases. However some variability among the assays have been noted and caution raised against O-glycans potentially masking antibody binding epitopes.sup.57. As POMC.sup.58, Kininogen-1.sup.59 and chromogranin A.sup.60 are also used as biomarkers it is particular important to note the degree of glycosylation identified in this study.
[0059] In summary the present inventors show that O-glycans in conserved residues in various peptide hormone families, are far more abundant than previously recognized, and that glycans change peptide hormone induced receptor activity and furthermore alter recognition by proteolytic enzymes that otherwise inactivate the peptide hormones.
EXAMPLES
[0060] The purpose of the following examples are given as an illustration of various embodiments of the invention and are thus not meant to limit the present invention in any way. Along with the present examples the methods described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Changes therein and other uses which are encompassed within the spirit of the invention as defined by the scope of the claims will occur to those skilled in the art.
[0061] Biological samples were prepared as follows:
[0062] Tissue Extraction: Porcine brain, cerebellum, ileum and a pool of human prostate gland tissue.sup.61 was isolated according to standard protocols. Proteins were extracted by crushing the frozen tissue using a CryoPrep tissue extractor (Covaris, Woburn, Mass.), boiled in water for 20 minutes, and homogenized with an Ultra-Turrax (IKA, Staufen, Germany). For ileum tissue, instead of boiling in water, the tissue was homogenized and rotated at 4° C. for 4 hours in 0.18 M HCl/70% ethanol. After 30 minutes centrifugation at 13,000 g, the supernatants were collected and pooled, and protein concentration was determined by BCA assay (Pierce). Prostate gland samples were further processed as crude water extract, acetone precipitated extract and acetone precipitated extract after acidification. Brain and cerebellum extracts were either crude extracts or acetone precipitated extracts after acidification. For precipitation of insoluble proteins samples in water or adjusted to 0.5 M CH3COOH were added icecold acetone (67%), incubated for 1 h at −20° C., and centrifuged at 16.000 g for 30 min. Subsequently the supernatant was lyophilized and reconstituted in water.
[0063] Plasma extraction: Plasma for O-glycoproteomic analyses was collected and pooled from two healthy volunteers into EDTA-treated tubes (K2E K2EDTA Vacuette) followed by centrifugation at 5,000×g for 10 min and stored at −20° C. until use.
[0064] Both crude and low molecular weight fraction (LMWF) enriched plasma was subjected to the O-glycoproteome strategy. For LMWF enrichment a volume of plasma containing approximately 60 mg of protein (measured by BCA assay (Pierce)) was precipitated by adding two parts of 96% ethanol followed by incubation at RT for 30 min. Supernatant and pellet was separated by centrifugation at 10.000×g for 10 minutes, and the supernatant was lyophilized and resuspended in 0.05% rapigest. 50 mM sodium acetate buffer and desialylated with 0.1 U/mL neuraminidase (Clostridium perfringens neuramidase type VI, Sigma). The LMWF-enriched sample was further enriched for O-glycopeptides by capture on a short (300 ml contained in 1 ml syringe) PNA agarose column. Glycoproteins were eluted by heating the lectin (2×90° C., 10 min) with 0.05% RapiGest (as previous described.sup.11). The LMWF-enriched plasma sample was 0.2 μm filtered prior to the glycoprotein enrichment. In parallel, 5 mg of total protein (as determined by BCA assay (Pierce)) from non-LMWF-enriched biofluid samples was desialylated by the same procedure as described above, before enzymatic degradation omitting the glycoprotein enrichment.
[0065] Cell protein extraction: Conditioned media cleared from dead cells and debris obtained from 2×T175 flasks cultured for 48-72 h were dialyzed, neuraminidase treated and enriched for glycoprotein as done for the the LMWF-enriched plasma. Total cell lysates (TCL) were obtained by washing a monolayer of cells in icecold PBS, scrabing off the cells and adding 2 ml 0.05% RapiGest to solubilize the cell pellet. The resulting homogenate was sonicated and cleared by centrifugation.
Mass Spectrometry Workflow
[0066] Enzyme digestion and desialylation: The extracted samples from the various neuronal and endocrine sources were adjusted to 50 mM ammonium bicarbonate, heated for 10 min at 80° C., followed by reduction with 5 mM dithiothreitol (DTT) (60° C., 30 min) and alkylation with 10 mM iodoacetamide (30 min, room temperature, kept dark). Subsequently, the samples were incubated with trypsin, Glu-C or chymotrypsin (Roche) (37° C., overnight, 1 μg enzyme pr 100 μg protein). The following day, the enzyme reaction was quenched and RapiGest, if present in the sample, was precipitated by acidifying with trifluoroacetic acid (TFA). The solution was cleared by centrifugation (10,000×g, 10 min.) and peptides were purified on C18 Sep-Pak columns (Waters), and dried down using SpeedVac. If not already desialylated, the dried peptides were resuspended in 1 mL 50 mM Sodium acetate (pH 5.5) containing 0.1 U/mL Neuraminidase followed by incubation at 37° C. for 1 h, purified by Sep-Pak and dried down.
[0067] LWAC O-glycopeptide enrichment: Dried samples were reconstituted in 2 mL PNA/Jacalin/VVA buffer (PNA-binding buffer 10 mM HEPES (pH 7.4), 150 mM NaCl, 0.1 mM CaCl.sub.2, and 0.01 mM MnCl.sub.2; Jacalin-binding buffer 175 mM Tris (pH 7.5); VVA-binding buffer 20 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM CaCl.sub.2/MgCl.sub.2/MnCl.sub.2/ZnCl.sub.2, and 1 M urea), 0.45 μm filtered and injected onto a pre-equilibrated 2.6-m long column packed with lectin-bound (PNA, Jacalin or VVA, Vector Laboratories) agarose beads at a constant flow-rate of 0.1 mL/min. For VVA the column was first washed for 3×CV in 0.4 M glucose and then eluted with 2 CV 0.2 M GalNAc and 1×CV 0.4 M GalNAc. For PNA and Jacalin LWAC, the column was washed 2×CV in lectin-binding buffer, and then eluted with 2×1 column volume 0.5 M galactose and 1×1 column volume 1 M galactose, respectively. The elution fractions were concentrated and glycopeptides were purified using Stage tips and submitted for nLCMS/MS analysis.
[0068] nLC/MS/MS Analysis: Liquid chromatography-tandem mass spectrometry was performed on a system composed of an EASY-nLC 1000 (Thermo Fisher Scientific) interfaced via a nanoSpray Flex ion source to an LTQ-Orbitrap Velos pro hybrid spectrometer or Orbitrap Fusion Tribrid (Thermo Fisher Scientific), equipped for both higher energy collisional dissociation (HCD) and electron transfer dissociation (ETD) modes, enabling peptide sequence analysis without and with retention of glycan site-specific fragments, respectively. The nLC was operated in a one-column set up with an analytical column (20 cm length, 75 μm inner diameter) packed with C18 reverse phase material (1.9-μm particle size, ReproSil-Pur, Dr. Maisch). Each sample dissolved in 0.1% formic acid was injected onto the column and eluted in a gradient from 2 to 30% B in 105 min, from 30% to 100% B in 5 min and 100% B for 10 min at 200 nl min-1 (solvent A, 100% H.sub.2O; solvent B, 100% acetonitrile; both containing 0.1% (v/v) formic acid). A data-dependent mass spectral acquisition routine, HCD triggering of subsequent ETD scan, was used for all runs. Briefly, a precursor MS1 scan (m/z 350-1,700) of intact peptides was acquired in the Orbitrap at a resolution setting of 30,000 (Velos Pro) or 120,000 (Fusion), followed by Orbitrap HCD-MS2 (m/z of 100-2,000) of the five most abundant multiply charged precursors in the MS1 spectrum; this event was followed up by an ETD-MS2 fragmentation for the same precursor ion. In cases where preliminary screening of fractions for glycopeptide enrichment was carried out prior to IEF, the ETD-MS2 step was omitted, and HCD-MS2 (m/z 100-2,000) of the five most abundant multiply charged precursors was acquired (“top five method”). These HCD-MS2 spectra were simply screened for the appearance of the HexNAc fragment at m/z 204.086.
[0069] Data analysis: The raw data were processed using Proteome Discoverer 1.4 software (Thermo Fisher Scientific) and searched against the human, porcine, mouse or rat-specific Uniprot database downloaded on January 2013. The Sequest HT search node was used for HCD and ETD data. In all cases the precursor mass tolerance was set to 15 p.p.m. and fragment ion mass tolerance to 20 millimass units (mmu). Carbamidomethylation on cysteine residues was used as a fixed modification. Methionine oxidation, C-terminal amidation (plasma and pancreas), and HexNAc or HexHexNAc attachment to serine, threonine or tyrosine were used as variable modifications. As an additional preprocessing procedure, all HCD data showing the presence of fragment ions at m/z 204.08 were extracted into a single .mgf file, and the exact mass of 1×, 2×, 3× and 4× HexNAc or HexHexNAc units was subtracted from the corresponding precursor ion mass, generating four distinct files. These preprocessed data files were submitted to a Sequest HT node under the same conditions mentioned above, except considering a HexNAc or HexHexNAc attachment. All spectra were searched against a decoy database using a target false discovery rate of 1%; unassigned spectra were submitted to a second Sequest HT node using the same parameters as above with the exception of performing the search using semi-specific trypsin cleavage. The final list was filtered to include only peptide hormones.
[0070] Multiple sequence alignment: All alignments were performed in ClustalW using the peptide sequences of H. sapiens, M. musculus, and R. Norvegicus.
[0071] In some aspects of the present application, mammalian host cells are modified to inactivate or downregulate certain glycosyltransferase genes. Details for for modifying or adding all subtypes of O-GalNAc linked mucin-type O-glycans are described in WO 2017/194699, which reference is hereby incorporated by reference.
[0072] In brief the present invention may incorporate the use of mammalian host cells with individual and combinatorial knock out of one or more of the GALNT1-T20 glycogenes (listed in Table 1 below). Determining changes in interactions with a plurality of mammalian cells with knock out of GALNT1 and/or GALNT2 and/or GALNT3 and/or GALNT4 and/or GALNT5 and/or GALNT6 and/or GALNT7 and/or GALNT9 and/or GALNT10 and/or GALNT11 and/or GALNT12 and/or GALNT13 and/or GALNT14 and/or GALNT16 and/or GALNT18 and/or GALNT19 is used to identify if said interaction occurs through subsets of O-GalNAc glycoproteins controlled by one or more of the 20 GALNTs, respectively, such that loss or reduction in measured interactions with mammalian cells with knock out of one or more of the named gene(s) confer that the O-glycoprotein(s) responsible for the interaction requires glycosylation by the corresponding GALNT(s).
TABLE-US-00001 TABLE 1 O-Gly (Ser/Thr/Tyr) GALNT1-T20 (O-GalNAc)
O-Glycan Branching
[0073] The present invention may incorporate the use of mammalian host cells with individual and combinatorial knock out of C1GalT1, GCNT1, GCNT2, GCNT3, GCNT4, GCNT6, GCNT7, B3GNT6 or B3GNT2 glycogenes (listed in Table 2 below) suitable for determining O-linked branching in Core2, Core3 and Core4 structures (
TABLE-US-00002 TABLE 2 O-Gly C1GALT1, B3GNT6 (O-GalNAc) GCNT1/T2/T3/T4/T6/T7 (O-GalNAc) B3GALNT1/T2 (O-Gly) B3GNT2/T3/T4/T7/T8/T9 (O-Gly) B4GALT1/T2/T3/T4 (O-Gly) B4GALNT3/T4 (O-Gly)
[0074] The present invention may incorporate the use of mammalian host cells with individual and combinatorial knock out of genes involved in N and O-glycan and glycolipid capping (sialylation); ST3GAL1/2/3/4/5/6 (α2,3NeuAc capping/sialylation) and/or ST6GAL1/2 (α2,6NeuAc capping/sialylation) and/or ST8SIA1/2/3/4/5/6 (capping by poly-sialylation) and/or ST6GALNAC1/2/3/4/5/6 (α2,6NeuAc capping/sialylation) (glycogenes listed in table 3 below) suitable for determining the capped (sialylated or fucosylated) glycan structure involved in observed interactions. Determining changes in interactions with a plurality of mammalian cells with knock out of ST3GAL1/2/3/4/5/6 and/or ST6GAL1/2 and/or ST8SIA1/2/3/4/5/6 and/or ST6GALNAC1/2/3/4/5/6 is used to identify if said interaction occurs through the type of capping indicated in parenthesis, such that loss or reduction in measured interactions with mammalian cells with knock out of one or more of the named groups of genes confer that the type of capping is responsible for the interaction as indicated.
TABLE-US-00003 TABLE 3 N-Gly, FUT1/2/3/4/5/6/7/8/9/10/11 O-Gly, ST3GAL1/2/3/4/5/6 Glycolipids ST6GAL1/2 ST6GALNAC1/2/3/4/5/6 ST8SIA1/2/3/4/5/6 B3GAT1/T2 ABO A4GNT
Engineering GTfs in Cells
[0075] Only little information exists as to the effects of knock out of glycosyltransferase genes in mammalian cell lines. For human cell lines only a few spontaneous mutants of glycosyltransferase genes have been identified. For example the colon cancer cell line LSC derived from LS174T has a mutation in the COSMC chaperone that leads to misfolded and non-functional Core1 synthase C1GalT.sup.62. The COSMC gene is also mutated in the human lymphoblastoid Jurkat cell line.sup.62,63.
Knock Out of Glycosylation Genes in Cell Lines
[0076] The limited information of effects of knock out of glycosyltransferase genes in cell lines is partly due to past difficulties with making knock outs in cell lines before the recent advent of precise gene editing technologies.sup.64. Thus, until recently essentially only one glycosyltransferase gene, FUT8, had been knocked out in a directed approach using two rounds of homologous recombination including massive clone screening efforts. The conventional gene disruption by homologous recombination is typically a very laborious process as evidenced by this knock out of Fut8 in CHO, as over 100,000 clonal cell lines were screened to identify a few growing Fut8−/− clones.sup.65 (U.S. Pat. No. 7,214,775). With the advent of the Zinc finger nuclease (ZFN) gene targeting strategy it became less laborious to disrupt genes, which was first demonstrated by knock out of the Fut8 gene in a CHO cell line, where additional two other genes unrelated to glycosylation were also effectively targeted.sup.66. More recently, TALENs and the CRISPR/Cas9 editing strategies have emerged, and the latter editing strategy was used to knock out the Fut8 gene.sup.67.
[0077] It is thus clear that targeted genetic engineering is now a tool the skilled person may use but editing of the glycosylation genes in mammalian cells and animals are prone to substantial uncertainty, and thus identifying the optimal engineering targets for display of a given glycan structure will require extensive experimental efforts. Therefore a random type approach involving testing of a multiplicity of different glycogene and glycoform variations may be beneficial.
Overexpression of Glycosylation Genes in Cell Lines
[0078] It is noteworthy that transient or stable overexpression of a glycosyltransferase gene in a cell most often result in only partial changes in the glycosylation pathways in which the encoded enzyme is involved. A number of studies have attempted to overexpress e.g. the core2 C2GnT1 enzyme in CHO to produce core2 branched O-glycans, the ST6GAL1 sialyltransferase to produce α2,6linked sialic acid capping on N-glycoproteins.sup.68, and the ST6GALNAC1 sialyltransferase to produce α2,6linked sialic acid on O-glycoproteins forming the cancer-associated glycan STn.sup.69. However, in all these studies heterogeneous and often unstable glycosylation characteristics in transfected cell lines have been obtained. This is presumably partly due to competing endogenous glycosyltransferase activities whether acting with the same substrates or diverging pathway substrates. Other factors may also explain the heterogeneous glycosylation characteristics.
Definitions
[0079] Before disclosing the subject-matter in greater detail, definitions of terms/expressions used herein are provided.
[0080] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
[0081] Each of the patents, applications and articles cited herein, and each document cited or referenced therein, including during the prosecution of any of the patents and/or applications cited herein (“patent cited documents”), and any manufacturer's instructions or catalogues for any products cited herein or mentioned in any of the references and in any of the patent cited documents, are hereby incorporated herein by reference. Documents incorporated by reference into this text or any teachings therein may be used in the practice of this invention.
[0082] Documents incorporated by reference into this text are not admitted to be prior art. As used herein, the words “may” and “may be” are to be interpreted in an open-ended, non-restrictive manner. At minimum, “may” and “may be” are to be interpreted as definitively including structure or acts recited.
[0083] The articles “a” and “an” are used herein to refer to one or to more than one (i.e. to at least one) of the grammatical object of the article.
[0084] Throughout this specification, unless the context requires otherwise, the words “comprise”, “comprises” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.
[0085] By “therapeutically effective amount or dose” or “sufficient amount or dose” herein is meant a dose that produces effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques.
[0086] The terms “pharmaceutically effective”, “therapeutically effective”, “pharmaceutically active”, or “therapeutically active” means that a synthetic compound of the invention so described is determined to have activity that affects a medical parameter or disease state.
[0087] “Patient” as that term is used herein, refers to the recipient of the treatment. In a specific embodiment, the patient is a mammal, such as a human, canine, murine, feline, bovine, ovine, swine or caprine. In a particular embodiment, the patient is a human.
[0088] As used herein, the terms “function” or “functional activity” refer to a biological, e.g., enzymatic function.
[0089] By “isolated” is meant material that is substantially or essentially free or purified from components that normally accompany it in its native state. For example, the compound according to the invention may be modified subsequent to isolation from their natural or laboratory-produced environment, or they may be used in isolated form in vitro, or as components of devices, compositions, etc.
[0090] By “obtained from” is meant that a sample such as, for example, a polypeptide (peptide hormone) is isolated from, or derived from, a particular source of the host or cells cultured in vitro. For example, the extract can be obtained from a tissue or a biological fluid sample isolated directly from the host. Therefore, the compounds of the present invention may be recombinantly produced or obtained from biological sources and be purified before further use in vitro and/or in vivo.
[0091] By “pharmaceutically acceptable carrier” is meant a solid or liquid filler, stabilizer, diluent or encapsulating substance that can be safely used in administration routes when applied to an animal, e.g. a mammal, including humans.
[0092] “Therapeutic treatment”, and “treatment”, refers any type of therapy.
[0093] The terms “peptide hormone”, “polypeptide,” “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply also to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.
[0094] The term “amino acid” refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid. Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.
[0095] As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a “conservatively modified variant” or “derivative” where the alteration results in the substitution of an amino acid, e.g., with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. According to the present invention, modified variants of the peptide hormones of the invention functionally retain their specific hormone activity when analyzed in a suitable model to determine said activity.
[0096] As used herein the term “peptide hormone” refers to any protein or peptide with hormonal activity, i.e. a peptide with signaling activity through the binding on its cognate receptor or to the class of peptide hormones involved in neuronal signaling, such as involved in a wide range of brain functions, including analgesia, reward, food intake, metabolism, reproduction, social behaviors, learning and memory. A peptide hormone may also be referred to as a neuropeptide.
[0097] The term “O-linked glycan” as used herein refers to the O-linked glycosylation with the addition of N-acetyl-galactosamine (GalNAc) to serine or threonine residues in the peptide hormones of the invention followed by other carbohydrates (such as galactose and sialic acid).
[0098] The phrase “at a predetermined specific site, such as in the receptor-binding region” as used herein refers to the addition or selection of peptides with an O-linked glycan at a site specifically selected.
[0099] The phrase “a truncated version or a variant as compared to the corresponding wild-type peptide hormone found in nature” is intended to refer to a peptide hormone which has been modified by either truncation or amino acid substitutions as compared with the same peptide hormone found in nature, such as found in vivo in the human body. Typically, the peptide hormone may be genetically engineered and/or chemically synthesized to include 1, 2, 3, 4, 5, 6, or 7 amino acids of the native peptide sequence being substituted with any other amino acid, such as conservative substitutions. Alternatively or in addition to this, 1, 2, 3, 4, 5, 6, or 7 amino acids may have been removed or added to the native peptide sequence. Accordingly, in some embodiments, the peptides hormone according to the present invention comprises 1, 2, 3, 4, 5, 6, or 7 substitutions, additions or deletions relative to the native wild-type peptide hormone. The amino acids used in the amino acid sequences according to the invention may be in both L- and/or D-form. It is to be understood that both L- and D-forms may be used for different amino acids within the same peptide sequence. In some embodiments the amino acids within the peptide sequence are in L-form, such as natural amino acids.
[0100] The term “improved stability” as used herein refers to peptides of the invention, which when tested e.g. in an in vitro assays as described in.sup.70 exhibit improved stability as compared to the same peptide without this one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region.
[0101] The term “receptor sub-type selectivity switch” as used herein refers to peptides of the invention being receptor sub-type specific.
[0102] The term “O-linked glycan” or “O-glycosylation” refers to the attachment of a sugar molecule to an oxygen atom in an amino acid residue in a protein.
Specific Embodiments of the Invention
[0103] 1. A formulation comprising at least one molecule of a peptide hormone species exhibiting a specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone species, wherein specific, defined glycosylation pattern means that the each molecule of said peptide hormone in said pharmaceutical formulation displays structural homogeneity with respect to the site of glycan attachment and/or with respect to the glycan attachment.
[0104] 2. A formulation comprising at least one molecule of a peptide hormone species according to embodiment 1, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 1, 2, 3, 5, 6, 7, 14, 15, 16, 21, 22, 23, 24, 25, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49, 51, 52, 54, 55, 56, 57, 72, 73, 74, 76, 79, 8, 81, 83, 84, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 106, 107, 108, 109, 113, 116, 117, 118, 119, 120, 130, 131, 135, 136, 143, 144, 147, 163, 164, 167, 168, 170, 184, 185, 186, 188, 189, 190, 191, 192, 204, 215, 217, 219, 222, 227, 229, 230, 231, 233, 234, 236, 252, 260, 262, 267, 272, and/or 279.
[0105] 3. A formulation comprising at least one molecule of a peptide hormone species according to embodiment 1 or 2, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 6, 7, 21, 22, 23, 24, 72, 74, 92, 95, 97, 99, 106, 107, 108, 116, 117, 118, 147, 163, 167, 185, 186, and/or 188.
[0106] 4. A formulation comprising at least one molecule of a peptide hormone species according to any one of embodiments 1 to 3, wherein said peptide hormone species is selected from the group of sequences comprising SEQ ID NOs: 72, 95, 97, 106, 108, 147, 185, and/or 188.
[0107] 5. A formulation comprising at least one molecule of a peptide hormone species according to any one of embodiments 1 to 4, wherein said peptide hormone species is SEQ ID NO: 147.
[0108] 6. A formulation comprising at least one molecule of a peptide hormone species according to any one of embodiments 1 to 5, wherein said predetermined specific site of said peptide hormone is within the receptor-binding region.
[0109] 7. A formulation comprising at least one molecule of a peptide hormone species according to any one of embodiments 1 to 6, wherein said peptide hormone exhibiting an specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone is obtainable by recombinant production using a host cell in which one or more glycosyltransferase gene(s) is/are inactivated or downregulated by inactivation and/or downregulation of one or more gene(s) selected from COSMC and C1GALT1; and/or by inactivation and/or downregulation of one or more gene(s) selected from GCNT3, GCNT4, B3GNT6, and/or by inactivation and/or downregulation and/or upregulation/activation of one or more gene(s) selected from ST6GALNAC1-6, ST3GAL1, GCNT3, GCNT4, and/or B3GNT6.
[0110] 8. A formulation comprising comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 7, wherein said peptide hormone species exhibits a specific glycosylation pattern of one or more O-linked glycan(s) at a predetermined specific site of said peptide hormone and, wherein the one or more O-glycan structures include a glycan structure selected from a Core1, core2, core3, or core4 structure with optional elongation and sialic acid capping, wherein optionally the first monosaccharide attached in the synthesis of O-linked glycans is N-acetyl-galactosamine and wherein a core1 structure may be obtained by the addition of galactose, and wherein a core2 structure may be obtained by the addition of N-acetyl-glucosamine to the N-acetyl-galactosamine of the core1 structure and wherein the core3 structures may be obtained by the addition of a single N-acetyl-glucosamine to the first monosaccharide N-acetyl-galactosamin and core4 structures may be obtained by the addition of a second N-acetyl-glucosamine to the core3 structure.
[0111] 9. A formulation comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 8, wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
[0112] 10. A formulation comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 9, wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
[0113] 11. A formulation comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 10, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
[0114] 12. A formulation comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 11, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
[0115] 13. A formulation comprising at least one molecule of a peptide hormone species according to any of embodiments 1 to 12, wherein the one or more O-glycan structures include the Core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
[0116] 14. A formulation according to any one of embodiments 1 to 13, wherein said formulation is a pharmaceutical formulation.
[0117] 15. A formulation comprising at least one molecule of a peptide hormone species as defined in any of the preceding embodiments 1 to 14, wherein said peptide hormone species comprises one or more O-linked glycan at a site indicated in and one of Tables 6A to 6E, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0118] 16. A formulation comprising at least one molecule of a peptide hormone species as defined in embodiment 15, wherein said peptide hormone species comprises one or more O-linked glycan at a site indicated in Table 6A or 6B, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0119] 17. A formulation comprising at least one molecule of a peptide hormone species as defined in embodiments 15 or 16, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6B, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0120] 18. A formulation comprising at least one molecule of a peptide hormone species as defined in any one of embodiments 15 to 17, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0121] 19. A formulation comprising at least one molecule of a peptide hormone species as defined in any one of embodiments 15 to 17, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0122] 20. A formulation comprising at least one molecule of a peptide hormone species as defined in any one of embodiments 15 to 17, wherein said peptide hormone species comprises one or more O-linked glycan at a site of a peptide hormone indicated in Table 6D, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone.
[0123] 21. A modified peptide hormone comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6A, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 1, 2, 3, 5, 6, 7, 14, 15, 16, 21, 22, 23, 24, 25, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49, 51, 52, 54, 55, 56, 57, 72, 73, 74, 76, 79, 8, 81, 83, 84, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 106, 107, 108, 109, 113, 116, 117, 118, 119, 120, 130, 131, 135, 136, 143, 144, 147, 163, 164, 167, 168, 170, 184, 185, 186, 188, 189, 190, 191, 192, 204, 215, 217, 219, 222, 227, 229, 230, 231, 233, 234, 236, 252, 260, 262, 267, 272, and/or 279.
[0124] 22. The modified peptide hormone according to embodiment 21 comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6B, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 6, 7, 21, 22, 23, 24, 72, 74, 92, 95, 97, 99, 106, 107, 108, 116, 117, 118, 147, 163, 167, 185, 186, and/or 188.
[0125] 23. The modified peptide hormone according to embodiments 21 or 22 comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6C, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is selected from the group comprising SEQ ID Nos: 72, 95, 97, 106, 108, 147, 185, and/or 188.
[0126] 24. The modified peptide hormone according to any one of embodiments 20 to 23 comprising one or more O-linked glycan at a predetermined specific site selected from the group comprising the peptide hormones indicated in 6D, particularly, wherein the site is a site in a human peptide hormone, more particularly, wherein the site is a conserved site in a human peptide hormone, and wherein said peptide hormone is depicted in SEQ ID NO: 147.
[0127] 25. The modified peptide hormone according to any one of embodiments 20 to 24, wherein the one or more O-glycan structures include a glycan structure selected from a Core1, core2, core3, or core4 structure with sialic acid capping, wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with optional elongation and sialic acid capping, wherein optionally the first monosaccharide attached in the synthesis of O-linked glycans is N-acetyl-galactosamine and wherein a core1 structure may be obtained by the addition of galactose, and wherein a core2 structure may be obtained by the addition of N-acetyl-glucosamine to the N-acetyl-galactosamine of the core1 structure and wherein the core3 structures may be obtained by the addition of a single N-acetyl-glucosamine to the first monosaccharide N-acetyl-galactosamin and core4 structures may be obtained by the addition of a second N-acetyl-glucosamine to the core3 structure.
[0128] 26. The modified peptide hormone according to any of embodiments 21 to 25, wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
[0129] 27. The modified peptide hormone according to any of embodiments 21 to 26, wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
[0130] 28. The modified peptide hormone according to any of embodiments 21 to 27, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
[0131] 29. The modified peptide hormone according to any of embodiments 21 to 28, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
[0132] 30. The modified peptide hormone according to any of embodiments 21 to 29, wherein the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
[0133] The present invention relates also to an isolated peptide hormone, such as recombinant, such as a neuropeptide comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region.
[0134] In some embodiments according to the present invention, the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with sialic acid capping, such as a structure as illustrated in
[0135] In some embodiments according to the present invention, the one or more O-glycan structures include a Tn (GalNAc) structure.
[0136] In some embodiments according to the present invention, the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
[0137] In some embodiments according to the present invention, the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
[0138] In some embodiments according to the present invention, the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
[0139] In some embodiments according to the present invention, the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
[0140] In some embodiments according to the present invention, the peptide hormone has improved, such as increased stability and/or circulatory half-life and/or other pharmacokinetic properties, such as improved stability in in vitro assays, plasma and/or bodyfluids.
[0141] In some embodiments according to the present invention, the peptide hormone has lower bioactivity in receptor signalling, such as decreased receptor stimulation in in vitro cell assays and/or in man.
[0142] In some embodiments according to the present invention, the peptide hormone exhibits improved receptor stimulation in in vitro cell assays and/or in animal models and/or in man.
[0143] In some embodiments according to the present invention, the peptide hormone exhibits altered blood-brain barrier uptake in animals or in man, such as increased blood-brain barrier uptake in animals or in man, or decreased blood-brain barrier uptake in animals or in human.
[0144] In some embodiments according to the present invention, the peptide hormone exhibits receptor sub-type selectivity switch.
[0145] In some embodiments according to the present invention, the peptide hormone is specific to one or more tissue in human, such as specific to tissue of the nervous system.
[0146] In some embodiments according to the present invention, the peptide hormone is selected from any one of tables 4, 5, or 6, such as selected from the list consisting of a peptide of the Neuropeptide Y family, such as NPY, PPY and PYY; a peptide of the Glucagon/Secretin family, such as GIP, Glucagon, GLP-1, GLP-2, PACAP, Secretin, PHM-27/PHV-42, Somatoliberin and VIP; a peptide of the Natriuretic peptide family, such as ANP, BNP and CNP, a peptide of the calcitonin family, such as calcitonin, and a peptide of the insulin family such as amylin.
[0147] In some embodiments according to the present invention, the peptide hormone is not found in nature. Accordingly, in some embodiments the peptide hormone according to the invention is not a wild-type hormone found in any species in nature. The peptide hormone according to the invention may be a peptide hormone that is a variant of a wild-type peptide hormone. Such peptide hormone variant may be a peptide that differ from the wild-type version by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid differences, such as amino acid substitutions, additions or deletions. A peptide variant according to the invention may have more than 80%, such as more than 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence identity with a corresponding wild-type peptide hormone found in nature.
[0148] In some embodiments according to the present invention, the peptide hormone is a truncated version or a variant as compared to the corresponding wild-type peptide hormone found in nature.
[0149] In some embodiments according to the present invention, the peptide hormone is selected from any one of table 6 comprising one or more O-linked glycan at a site as indicated in table 6, such as at a bold underlined position and/or an italic underlined position.
[0150] In some embodiments according to the present invention, the peptide hormone is selected from any one of table 6 comprising at least, not more than, or the exact number of O-linked glycan sites as indicated in table 6.
[0151] In some embodiments according to the present invention, the peptide hormone is selected from any one of table 5.
[0152] Subject matter of the present invention is also a (pharmaceutical or diagnostic) composition or formulation comprising a compound as defined in any of the preceding embodiments.
[0153] Subject matter of the present invention is also a (pharmaceutical or diagnostic) composition or formulation comprising a synthetic compound as defined in any of the preceding embodiments, wherein said (pharmaceutical or diagnostic) composition or formulation is suitable for administration to a patient in need thereof.
[0154] Subject matter of the present invention is also a peptide hormone or a pharmaceutical composition or formulation according to any of the above embodiments, wherein said composition or formulation is suitable for the localized or systemic administration, wherein the localized administration is preferably selected from the group of topical administration, including transdermal, ophthalmic, nasal, otologic, enteral, pulmonal and urogenital administration or local or systemic injection, including subcutaneous, intra-articular, intravenous, intracardiac, intramuscular, intraosseous or intraperitoneal administration.
[0155] Subject matter of the present invention is also a device according to the previous embodiment, wherein the device is suitable as a delivery system for immediate and/or sustained release of a peptide hormone as defined in anyone of the preceding embodiments, e.g., they may be used as drug (peptide hormone) delivery system or controlled drug (peptide hormone) release systems.
[0156] Subject-matter of the invention is also a device comprising a pharmaceutical composition or a pharmaceutical formulation as defined in any of the foregoing embodiments. A device may take any form that is suitable to deliver the synthetic compounds or any one of the compositions or formulations of the present invention. It may comprise biological and/or synthetic materials and may take form of a patch, a stent, an implantable device, hydrogel, etc.
[0157] Subject-matter of the invention is also a device as defined in any of the foregoing embodiments, wherein the device is a delivery system for immediate and/or sustained release of the peptide hormone as defined in any of the foregoing embodiments.
[0158] Subject-matter of the invention is also a method of treatment of an individual in need thereof and/or the amelioration of and/or the prevention of deterioration of a disease in an individual in need thereof, by administration to said individual of a therapeutically efficient amount of any of the peptide hormones according to the present invention and/or pharmaceutical compositions as defined above.
[0159] The administration of the compounds (peptide hormones) according to this invention and pharmaceutical compositions according to the invention may be performed in any of the generally accepted modes of administration available in the art. Illustrative examples of suitable modes of administration include intravenous, oral, nasal, inhalable, parenteral, topical, transdermal and rectal delivery. Parenteral and intravenous delivery forms are preferred. In aspects of the invention injectable formulations comprising a therapeutically effective amount of the compounds (peptide hormones) of the invention are provided, including salts, esters, isomers, solvates, hydrates and polymorphs thereof, at least one vehicle comprising water, aqueous solvents, organic solvents, hydro-alcoholic solvents, oily substances, or mixtures thereof, and optionally one or more pharmaceutically acceptable excipients. Standard knowledge regarding these pharmaceutical ingredients and pharmaceutical formulations/compositions may be found, inter alia, in the ‘Handbook of Pharmaceutical Excipients’; Edited by Raymond C Rowe, Paul J Sheskey, Walter G Cook and Marian E Fenton; May 2012 and/or in Remington: The Science and Practice of Pharmacy, 19th edition. The pharmaceutical compositions/formulations may be formulated in the form of a dosage form fororal, intravenous, nasal, inhalable, parenteral, topical, transdermal and rectal and may thus comprise further pharmaceutically acceptable excipients, such as buffers, solvents, preservatives, disintegrants, stabilizers, carriers, diluents, fillers, binders, lubricants, glidants, colorants, pigments, taste masking agents, sweeteners, flavorants, plasticizers, and any acceptable auxiliary substances such as absorption enhancers, penetration enhancers, surfactants, co-surfactants, and specialized oils.
[0160] The proper excipient(s) is (are) selected based in part on the dosage form, the intended mode of administration, the intended release rate, and manufacturing reliability. Examples of common types of excipients include also various polymers, waxes, calcium phosphates, sugars, etc.
[0161] Polymers include cellulose and cellulose derivatives such as HPMC, hydroxypropyl cellulose, hydroxyethyl cellulose, microcrystalline cellulose, carboxymethylcellulose, sodium carboxymethylcellulose, calcium carboxymethylcellulose, and ethylcellulose; polyvinylpyrrolidones; polyethylenoxides; polyalkylene glycols such as polyethylene glycol and polypropylene glycol; and polyacrylic acids including their copolymers and crosslinked polymers thereof, e.g., Eudragit® (Rohm), polycarbophil, and chitosan polymers. Waxes include white beeswax, microcrystalline wax, carnauba wax, hydrogenated castor oil, glyceryl behenate, glycerylpalmitol stearate, and saturated polyglycolyzed glycerate. Calcium phosphates include dibasic calcium phosphate, anhydrous dibasic calcium phosphate, and tribasic calcium phosphate. Sugars include simple sugars, such as lactose, maltose, mannitol, fructose, sorbitol, saccharose, xylitol, isomaltose, and glucose, as well as complex sugars (polysaccharides), such as maltodextrin, amylodextrin, starches, and modified starches.
[0162] The pharmaceutical compositions/formulations of the present invention may be formulated into various types of dosage forms, for instance as solutions or suspensions, or as tablets, capsules, granules, pellets or sachets for oral administration.
[0163] The pharmaceutical composition of the present invention can be manufactured according to standard methods known in the art. Granulates according to the invention can be obtained by dry compaction or wet granulation. These granulates can subsequently be mixed with e.g. suitable disintegrating agents, glidants and lubricants and the mixture can be compressed into tablets or filled into sachets or capsules of suitable size. Tablets can also be obtained by direct compression of a suitable powder mixture, i.e. without any preceding granulation of the excipients. Suitable powder or granulate mixtures according to the invention are also obtainable by spray drying, lyophilization, melt extrusion, pellet layering, coating of the active pharmaceutical ingredient or any other suitable method. The so obtained powders or granulates can be mixed with one or more suitable ingredients and the resulting mixtures can be delivered in sterile primary packaging devices for reconstitution before parenteral administration Injectable compositions of the present invention may contain a buffer (for example, sodium dihydrogen phosphate, disodium hydrogen phosphate and the like), an isotonizing agent (for example, glucose, sodium chloride and the like), a stabilizer (for example, sodium hydrogen sulfite and the like), a soothing agent (for example, glucose, benzyl alcohol, mepivacaine hydrochloride, xylocaine hydrochloride, procaine hydrochloride, carbocaine hydrochloride and the like), a preservative (for example, p-oxybenzoic acid ester such as methyl p-oxybenzoate and the like, thimerosal, chlorobutanol, benzyl alcohol and the like) and the like, if necessary. In addition, the injectable composition of the present invention may contain vitamins and the like. Further, injectable compositions of the present invention may contain an aqueous solvent, if necessary. Examples of the aqueous solvent include purified water for injection, physiological saline solution, and glucose solution. In injectable compositions of the present invention, the pharmaceutical compound (peptide hormone) may be solid. As used herein, the “solid” comprises crystals and amorphous substances which have conventional meanings. The form of the solid component is not particularly limited, but powder is preferred in view of dissolution rate.
Pharmaceutical Formulations
[0164] Still another aspect of the present invention relates to the use of the peptide hormones according to the present invention, e.g. as shown in Tables 6A-E) as an active ingredient, together with at least one pharmaceutically acceptable carrier, excipient and/or diluents for the manufacture of a pharmaceutical composition for the treatment and/or prophylaxis of appropriate disorders or diseases.
[0165] Administration forms include, for example, pills, tablets, film tablets, coated tablets, capsules, liposomal formulations, micro- and nano-formulations, powders and deposits. Furthermore, the present invention also includes pharmaceutical preparations for parenteral application, including dermal, intradermal, intragastral, intracutan, intravasal, intravenous, intramuscular, intraperitoneal, intranasal, intravaginal, intrabuccal, percutan, rectal, subcutaneous, sublingual, topical, or transdermal application, which preparations in addition to typical vehicles and/or diluents contain the compounds according to the present invention.
[0166] The compounds of the invention can also be administered in form of its pharmaceutically active salts. Suitable pharmaceutically active salts comprise acid addition salts and alkali or earth alkali salts. For instance, sodium, potassium, lithium, magnesium or calcium salts can be obtained.
[0167] The pharmaceutical compositions/formulations according to the present invention will typically be administered together with suitable carrier materials selected with respect to the intended form of administration, i.e. for oral administration in the form of tablets, capsules (either solid filled, semi-solid filled or liquid filled), powders for constitution, aerosol preparations consistent with conventional pharmaceutical practices. Other suitable formulations are hydrogels, elixirs, dispersible granules, syrups, suspensions, creams, lotions, solutions, emulsions, suspensions, dispersions, and the like. Suitable dosage forms for sustained release include tablets having layers of varying disintegration rates or controlled release polymeric matrices delivered with the active components. The pharmaceutical compositions may be comprised of 0.01 to 95% by weight of the peptide hormones of the invention.
[0168] As pharmaceutically acceptable carrier, excipient and/or diluents can be used HSA, lactose, sucrose, cellulose, mannitol.
[0169] Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethyl-cellulose, polyethylene glycol and waxes. Among the lubricants that may be mentioned for use in these dosage forms, boric acid, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include starch, methylcellulose, guar gum and the like. Sweetening and flavoring agents and preservatives may also be included where appropriate. Some of the terms noted above, namely disintegrants, diluents, lubricants, binders and the like, are discussed in more detail below. Additionally, the compositions of the present invention may be formulated in sustained release form to provide the rate controlled release of any one or more of the components or active ingredients to optimize the therapeutic effects. Suitable dosage forms for sustained release include controlled release polymeric matrices or hydrogels embedding the active components. Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier such as inert compressed gas, e.g. nitrogen.
[0170] For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides such as cocoa butter is first melted, and the active ingredient is dispersed homogeneously therein by stirring or similar mixing. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.
[0171] Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for parenteral administration. Such liquid forms include solutions, suspensions and emulsions.
[0172] The compounds of the present invention may also be deliverable transdermally. The transdermal compositions may take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.
[0173] The transdermal formulation of the compounds of the invention is understood to increase the bioavailability of said compound into the circulating blood. One problem in the administration of peptidic drugs in general is the loss of bioactivity due to the formation of insolubles in aqueous environments or due to degradation. Therefore, stabilization of compounds for maintaining their fluidity and maintaining their biological activity upon administration to the patients in need thereof needs to be achieved. Prior efforts to provide active agents for medication include incorporating the medication in a polymeric matrix whereby the active ingredient is released into the systemic circulation. Known sustained-release delivery means of active agents are disclosed, for example, in U.S. Pat. Nos. 4,235,988, 4,188,373, 4,100,271, US447471, U.S. Pat. Nos. 4,474,752, 4,474,753, or U.S. Pat. No. 4,478,822 relating to polymeric pharmaceutical vehicles for delivery of pharmaceutically active chemical materials to mucous membranes. The pharmaceutical carriers are aqueous solutions of certain polyoxyethylene-polyoxypropylene condensates. These polymeric pharmaceutical vehicles are described as providing for increased drug absorption by the mucous membrane and prolonged drug action by a factor of two or more. The substituents are block copolymers of polyoxypropylene and polyoxyethylene used for stabilization of drugs such as insulin.
[0174] Aqueous solutions of polyoxyethylene-polyoxypropylene block copolymers (poloxamers) are useful as stabilizers for the compounds. Aside from serving as a stabilizer for the compound, poloxamers provide excellent vehicles for the delivery of the compound, and they are physiologically acceptable. Poloxamers, also known by the trade name Pluronics (e.g. Pluronic F127, Pluronic P85, Pluronic F68) have surfactant properties that make them useful in industrial applications. Among other things, they can be used to increase the water solubility of hydrophobic, oily substances or otherwise increase the miscibility of two substances with different hydrophobicities. For this reason, these polymers are commonly used in industrial applications, cosmetics, and pharmaceuticals. They have also been used as model systems for drug delivery applications. In situ gelation of pharmaceutical compositions based on poloxamer that are biologically triggered are known in the art (e.g. U.S. Pat. No. 5,256,396), describing compositions containing poloxamer 407 and water at specified concentrations.
[0175] Gels refer to the active ingredients dispersed or solubilized in a hydrophilic semi-solid matrix. Powders for constitution refer to powder blends containing the active ingredients and suitable diluents which can be suspended in water and may contain optionally buffer salts, lactose, amino acids, excipients, sugars and isotonisation reagents.
[0176] Recently, increasingly improved and potent protein-based and peptide-based drugs have been developed by the biotech industry. However, the prophylactic and/or therapeutic use of many other protein- or peptide-based compounds has been hampered because of their susceptibility to proteolytic breakdown, rapid plasma clearance, peculiar dose-response curves, immunogenicity, bioincompatibility, and/or the tendency of peptides and proteins to undergo aggregation, adsorption, and/or denaturation. These characteristics often render traditional methods of drug delivery ineffective or sub-optimal when applied to protein or peptide based drugs. Therefore, an immense amount of interest has been increasingly placed on controlled and/or sustained release drug delivery systems to maintain the therapeutic efficacy or diagnostic value of these important classes of biologically active agents. One of the primary goals of sustained delivery systems is to maintain the levels of an active agent within an effective range and ideally at a constant level. One approach for sustained delivery of an active agent is by microencapsulation, in which the active agent is enclosed within a polymeric matrix. The importance of biocompatible and/or biodegradable polymers as carriers for parenteral drug delivery systems is now well established. Biocompatible, biodegradable, and relatively inert substances such as poly(lactide) (PLA) or poly(lactide-co-glycolide) (PLG) structures such as microparticles or films containing the active agent to be administered are commonly employed sustained delivery devices (for review, see M. Chasin, Biodegradable polymers for controlled drug delivery. In: J.O. Hollinger Editor, Biomedical Applications of Synthetic Biodegradable Polymers CRC, Boca Raton, Fla. (1995), pp. 1-15; T. Hayashi, Biodegradable polymers for biomedical uses. Prog. Polym. Sci. 19 4 (1994), pp. 663-700; and Harjit Tamber, Pal Johansen, Hans P. Merkle and Bruno Gander, Formulation aspects of biodegradable polymeric microspheres for antigen delivery Advanced Drug Delivery Reviews, Volume 57, Issue 3, 10 Jan. 2005, Pages 357-376). A relatively steady release of one or more active agents incorporated within such polymers is possible because of the degradation profile of these polymers in an aqueous environment. By encapsulating active agents in a polymer matrix in various forms such as microparticles and/or films the active agent is released at a relatively slow rate over a prolonged time. Achieving sustained drug release in such a manner may afford less frequent administration, thereby increasing patient compliance and reducing discomfort; protection of the therapeutic compound within the body; potentially optimized prophylactic or therapeutic responses and prolonged efficacy; and avoidance of peak-related side-effects by maintaining more-constant blood levels of the active agent. Furthermore, these compositions can oftentimes be administered by injection, allowing for localized delivery and high local concentrations of the active agents.
[0177] With regard to highly active biologies, such as growth factors, local in the form of a bolus injection results in rapid diffusion from the region of interest and can cause severe side effects and limit efficacy. The oldest way is to use biophysical retention by changing the biophysical properties in form of viscosity, porosity, hydrophobicity or charge of the material to attain a purposeful delivery. This strategy often substantially modifies the properties of the tissue and conditions for cells, requiring more appropriate, biocompatible release mechanisms.
Methods of Treatment
[0178] Treatment methods of the invention comprise the step of administering to a subject a therapeutically effective amount of at least one peptide hormone according to the invention or a pharmaceutical composition/formulation of the invention. The administration may be effected by any route, e.g., dermally, parenterally, topically, etc.
[0179] As indicated previously “therapeutically effective amount” of a at least one peptide hormone according to the invention preferably refers to the amount necessary to achieve the therapeutic outcome.
[0180] The choice of the optimal dosage regime and duration of medication, particularly the optimal dose and manner of administration of the active compounds necessary in each case can be determined by a person skilled in the art on the basis of his/her expert knowledge.
[0181] Subject matter of the present invention is also any of the above the above at least one peptide hormone in method of manufacturing a medicament for the treatment of an appropriate condition or diseases.
EXAMPLES
Example 1
Development of a Sensitive O-Glycoproteomics Work Flow Enriching for Peptide Hormones
[0182] The present inventors originally developed the so-called SimpleCell O-glycoproteomics strategy.sup.18,71, and provided a vast expansion of the knowledge of the human O-glycoproteome. While more than 3,000 O-glycosites were identified in almost 1,000 human proteins.sup.28,71. In order to specifically explore potential O-glycans on peptide hormones the present inventors developed a novel proteomics based strategy selective for smaller peptides. The overall strategy for exploring O-glycosylation of peptide hormones is presented in
[0183] Peptide hormones are typically short peptides of 30-50 amino acids, and to achieve optimal peptide hormone coverage in the mass spectrometry analysis, the present inventors used different pre-extraction methods for the tissue- and plasma samples (precipitation with organic solvents or acidic water extraction) to ensure enrichment of shorter peptides. For the cell lines, the present inventors furthermore used different proteolytic enzymes (trypsin, Glu-C and/or chymotrypsin) for protein digestion to facilitate better coverage of proteins in the LWAC-LC/MS workflow (
[0184] When studying protein O-glycosylation in complex mixtures enrichment of glycopeptides is essential for glycan detection due to suppression of glycopeptide ionization by the presence of large amounts of unglycosylated peptides.sup.72. In the case of N2A cells expressing truncated O-glycans due to a spontaneous mutation in the COSMC gene.sup.73, the Tn (GalNAc) binding VVA (vicia villosa lectin) was used as previously described. Normal cells generally produce elongated O-glycans of core 1 or in some cases cores 2-4 structures with capping sialic acids (
Example 2
Identification of O-Glycans on Peptide Hormones
[0185] In total the present inventors identified 2,327 O-glycoproteins (199 from human, 898 from pig, 509 from rat and 721 from mouse) with approximately 5,000 unambiguously assigned sites and another approximately 8,000 ambiguously assigned modification specific O-glycopeptides where site information was not obtained due to poor quality spectra or lack of ETD data from 10 different tissues and multiple species (
[0186] In order to extract information on glycosylated peptide hormones across several species (see
[0187] To further explore the total number of O-glycosylated peptide hormones across the mammalian species analysed, the present inventors realigned the resulting peptide fragments to the corresponding human homolog resulting in 62 preproproteins (
[0188] On average, our strategy resulted in the identification of 10-20 glycosylated peptide hormones per analyzed sample, with the exception of the plasma and brain samples that resulted in identification of 5 and 39 glycosylated peptide hormones, respectively (
Example 3
O-Glycans Identified in Conserved Receptor Binding Domains of Peptide Hormone Families
[0189] PTMs are known to change the biochemical properties and diversify protein function. In particular O-glycosylation in close proximity to limited proteolytic cleavage sites has been demonstrated to e.g. co-regulate limited proteolytic processing. Therefore, the present inventors first explored if the identified sites were in close proximity (+/−3 a.a.) to physiological relevant cleavage sites in peptide hormones. However, mapping the identified sites relative to peptide hormone length revealed that this was not the case.
[0190] The major characterized structural or functional features of peptide hormones, besides limited proteolytic processing, are their ability to be recognized by and bind highly selective receptors. The present inventors therefore explored the positions of glycosites relative to characterized structural and/or functional receptor binding regions, and surprisingly, in six different peptide hormone families the majority of identified or predicted sites were indeed located in known receptor binding regions (
[0191] The following details findings in each of the hormone families. Glycosylated residues are numbered according to the length of the mature processed bioactive peptide hormones:
Example 4
O-Glycans Identified in Conserved Receptor Binding Domains of the Secretin/Glucagon Family
[0192] The members of the Secretin/Glucagon family function in water homeostasis and regulation of feeding behavior and have remarkable sequence homology. Here the present inventors identified O-glycans on Secretin, Vasoactive Intestinal Peptide (VIP), Peptide Histidine Methionine/valine (PHM-27/PHV-42), Glucagon and Glucagon-like peptide 1(GLP-1), positioned in the N-terminal part of the peptide hormones, which has been shown to be important for receptor binding and activation.sup.80 (
Example 5
O-Glycans Identified in Conserved Receptor Binding Domains of the Calcitonin Family
[0193] Members of the Calcitonin control a number of processes, including calcium/phosphate balance (Calcitonin), insulin dependent glucose metabolism (Amylin/IAPP) and vasodilation (Adrenomedullin and Intermedin).sup.81. The present inventors identified O-glycans on all members of the Calcitonin family with the exception of Intermedin (
Example 6
O-Glycans Identified in Conserved Receptor Binding Domains of the Insulin-Like Growth Factor Family
[0194] In the IGF/Insulin subfamily of the Insulin gene superfamily, the present inventors identified an O-glycan on Insulin in the B-chain at Thr27 in a semi-conserved residue found as a serine in Insulin Growth Factor II (IGFII) and a threonine shifted a few positions C-terminally in Insulin Growth Factor I (IGFI) (
Example 7
O-Glycans Identified in Conserved Receptor Binding Domains of the Galanin Family
[0195] Galanin and Galanin-like peptide have multiple functions stimulating smooth muscle cell contraction and growth hormone and insulin release. On Galanin itself the present inventors identified an O-glycan on Thr3, which is an essential residue for receptor activation.sup.84 and conserved in both members. In addition to this the present inventors found an O-glycan on Ser11, which is only present in Galanin and not the other family member Galanin-like peptide (
Example 8
O-Glycans Identified in Conserved Receptor Binding Domains of the Neuropeptide Y Family
[0196] The Neuropeptide Y family members, Neuropeptide Y (NPY), Peptide YY (PYY) and Pancreatic Polypeptide (PPY) share structural features and they all adopt a specific three-dimensional structure called the PP-fold. The peptides are involved in appetite regulation and anxious behavior, and the present inventors found all three members to carry O-glycans at the same conserved C-terminal Thr32 residue (
Example 9
O-Glycans Identified in Conserved Receptor Binding Domains of the Natriuretic Peptides Family
[0197] The present inventors identified O-glycans on all three precursors (pro Atrial Natriuretic peptide (ANP), pro B-type Natriuretic Peptide (BNP), pro C-type Natriuretic Peptide (CNP)) as previously described for proBNP.sup.79. However, the present inventors also identified O-glycans at Ser19 and Ser25 in the C-terminal cyclic receptor-binding region of mature ANP where Ser19 is highly conserved in all three members (
O-glycans on peptide hormones modulate receptor activation.
[0198] Considering the high degree of conservation and structural position of the identified O-glycans the present inventors next decided to explore the potential functional impact of site-specific O-glycosylation on mature peptide hormones. The present inventors selected Glucagon, GLP-1, Secretin, VIP, ANP, NPY, PYY and PPY as examples of peptide hormones where the present inventors identified O-glycosylation sites in known receptor activating regions for analysis in in vitro receptor binding assays. First, the present inventors used recombinant GalNAc-transferases for chemoenzymatic synthesis or chemically synthesized (SynPeptides, China) glycopeptide variants with Tn (GalNAcα1-O-Ser/Thr), elongated to T (Galβ1-3GalNAcα1-O-Ser/Thr) and sialylated ST (NeuAcα2-3Galβ1-3GalNAcα1-O-Ser/Thr) structures using recombinant purified glycosyl transferases. Secretin, Glucagon. GLP-1 and VIP were enzymatically GalNAc-glycosylated by GalNAc-T1 corresponding to position Thr7 (MS validated) in the mature peptide sequences and peptides from the NPY family were chemically synthesized with GalNAc at position Thr32. ANP was chemically synthesized with an O-glycan at position 19 and/or 25. Next HEK293 or COS-7 cells transiently expressing selected relevant cognate receptors were incubated with increasing concentrations of peptide or glycopeptide. Ligand/agonist efficacy and potency was measured by receptor activation as an increase in secondary messenger cAMP.
[0199] Cell Culture and Transfection: HEK293 and COS7-cells were cultured in DMEM containing 10% FBS in a humidified atmosphere at 37° C. with 5% C02 (Sigma-Aldritch, Germany). For experiments, cells were seeded onto 6- or 10-cm plates and cultured for 1-3 days to 60-80% confluency. Cells were transfected with 1-2.5 pg of receptor constructs for 24-48 h using Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer's instructions, or alternatively, using linear 25-kDa polyethyleneimine (Polysciences). Both reagents were used at 1:3 DNA to reagent ratio.
[0200] cAMP and cGMP Accumulation Measured by HTRF®: Intracellular cAMP/cGMP levels were measured using a homogeneous time-resolved fluorescence (HTRF®) cAMP/cGMP Gs dynamic assay kit (CisBio Bioassays). Forty-eight hours post-transfection cells were detached and seeded into white 384-well microplates with 1,000 cells/well in 5 μl of stimulation buffer (DMEM, 1 mM 3-isobutyl-1-methylxanthine (IBMX), 0.2% BSA). For their stimulation, μl/well of the stimulation buffer containing appropriate doses of either naked peptide hormone or glycosylated variants were added. Then, cells were incubated for 30 min at 37° C. followed by lysis by addition of 5 μl/well of each of the supplied conjugate-lysis buffer containing d2-labeled cAMP/cGMP and Europium cryptate-labeled anti-cAMP/cGMP antibody, both reconstituted according to the manufacturer's instructions. Plates were incubated for 1 h in the dark at room temperature and time-resolved fluorescence signals were excited at 340 nm and measured at 620 and 665 nm, respectively using the EnSpire Multilabel Reader (PerkinElmer Life and Analytical Sciences). The cAMP/cGMP generated was interpolated from a cAMP standard curve generated in parallel for each experiment.
[0201] IP-1 Accumulation Measured by HTRF®: Intracellular IP-1 levels were measured, similarly to cAMP, using a homogeneous time-resolved fluorescence (HTRF®) IP-1 Gq assay kit (CisBio Bioassays). This assay is dependent on co-transfection of the receptor with a chimeric G-protein to obtain sufficient signal of the Gq pathway. In this assay NPY2R was used together with Gqo5. Forty-eight hours post-transfection cells were detached and seeded into white 384-well microplates with 10,000 cells/well in 7 μl of supplied stimulation buffer supplemented with 0.1% BSA. For stimulation, 7 μl/well of the supplemented stimulation buffer containing appropriate doses of either naked peptide hormone or glycosylated variants were added. Then, cells were incubated for 2 h at 37° C. followed by lysis by addition of 3 μl/well of each of the supplied conjugate-lysis buffer containing d2-labeled IP-1 and Europium cryptate-labeled anti-IP-1 antibody, both reconstituted according to the manufacturer's instructions. Plates were incubated for 1 h in the dark at room temperature and time-resolved fluorescence signals were excited at 340 nm and measured at 620 and 665 nm, respectively using the EnSpire Multilabel Reader (PerkinElmer Life and Analytical Sciences). The IP-1 generated was interpolated from a IP-1 standard curve generated in parallel for each experiment.
Example 10
O-Glycans on VIP Modulate Receptor Activation.
[0202] The two VIP/PACAP receptors (VPAC1 and VPAC2) show comparable affinities for VIP.sup.52,86, thus the present inventors selected VPAC1 for analysis of VIP binding and activation. In this assay VIP exhibited a potency of 0.2 nM and 0.4 for VPAC1 & 2 respectively, which is in good agreement with previous studies. VIP with one O-glycan (GalNAc/Tn) at residue 7 (VIP-Thr7/Tn) showed a 581-fold decrease in potency to 102 nM for VPAC1 (
Example 11
O-Glycans on Secretin Modulate Receptor Activation.
[0203] Secretin binds and signals exclusively through the secretin receptor (SCTR), and in our assay secretin had a potency of 0.1 nM for SCTR, comparable to values found in previous studies, whereas secretin with a single O-glycan (GalNAc/Tn) at residue 7 (Secretin-Thr7-Tn) decreased the potency 2200-fold to 205 nM. Elongation of the glycan on secretin to T and ST structures further reduced potency 1-7 fold for each elongation step to 292 nM and 1932 nM, respectively (
Example 12
O-Glycans on GLP-1 Modulates Receptor Activation.
[0204] GLP-1 binds and signals exclusively through the GLP-1 receptor (GLP-1R), and in our assay GLP-1 showed a potency of 0.04 nM for GLP-1R, comparable to values found in previous studies. Elongation of the glycan at position 7 on GLP-1 to T, ST and diST (GLP1-Thr7/Tn/T/diST) further reduced potency approximately 20-40 fold fold for each elongation step to 253 nM, 266 nM, and 461 nM, respectively (
Example 13
O-Glycans on Glucagon Modulates Receptor Activation.
[0205] As with GLP-1 and the GLP-1R, Glucagon binds and signals exclusively through the glucagon receptor (GCGR). The non-glycosylated glucagon showed a potency of 1.29 nM in line with previous literature. Upon introducing a Tn-glycoform at Thr7 (Glucagon-Thr7/Tn), the potency is decreased almost a 100-fold to 126 nM. This potency is reduced 5 times further when elongating to T (667.9 nM). However, the introduction of sialic acid (ST) did not significantly influence the potency further compared to the non-sialylated T-structure (615.7 nM). Importantly, removal of the glycan from the T-glycosylated glucagon restored the potency to the non-glycosylated levels eliciting an EC50 of 1,235 nM (
Example 14
O-Glycans on NPY, PYY and PPY Modulate Receptor Activation.
[0206] The NPY family peptide hormones activate members of the NPY receptor family (Y1, Y2, Y4, and Y5), where mature NPY (1-36) and PYY (1-36) preferentially binds Y1, Y2 and Y5, and PPY preferentially binds Y4. The Y1 receptor seem to have strict requirements for the N-terminal part of the peptides as N-terminal truncation gradually decreases affinity for NPY. In contrast the Y2 receptor is more sensitive to alterations in the C-terminus of NPY and PYY and single amino acid substitutions in the C-terminus can lower affinity for the Y2 receptor.sup.87.
[0207] The present inventors developed a receptor assay for the binding study of the NPY family to the four known receptors. As expected according to reference values in the literature PYY, NPY, had respective potencies (EC50-values) of 0.47 nM and 2.16 nM at NPY1R, 0.34 nM and 4.11 nM at receptor NPY2R, 11.35 nM and 199.9 nM at receptor NPY4R, and 12.04 and 15.03 nM at receptor NPY5R (
Example 15
O-Glycans on ANP Modulate Receptor Activation.
[0208] ANP exerts its physiological effects mainly via the NPR-A receptor but binds also to NPR-C. NPR-C, however, is mainly regarded a clearance receptor thus the present inventors selected NPR-A for ANP binding and activation.
[0209] In this assay ANP exhibited a potency of 0.9 nM for NPR-A, which is in good agreement with previous studies.sup.88. ANP with one O-glycan (GalNAc/Tn) at residue 19 or 25 showed a 63- and 139-fold decrease in potency to 57 and 125 nM, respectively, whereas simultaneous O-glycans at residue 19 and 25 completely dismiss receptor activation. Elongation of the O-glycan on ANP residue 19 to T, ST and diST further reduced the potency 3- to 50-fold to 160 nM and 296 nM and 2699 nM, respectively. Elongation of the O-glycan on ANP residue 25 to T, ST and diST further reduced the potency 2- to 3-fold to 265 nM and 717 nM and 460 nM respectively (
[0210] In summary, all peptide hormones with O-glycans attached at one or more specific sites elicited a right-shifted full, partial agonist or superagonist response positively correlated to glycan size.
Example 16
O-Glycans Modulate Peptide Hormone Stability In Vitro
[0211] Many peptide hormones are destined for endocrine circulation where they are rapidly degraded with half-lives reaching only a few minutes.sup.6-8. The present inventors have previously demonstrated that O-glycans in close proximity to proteolytic processing sites can modulate the rate of processing.sup.33. To study if O-glycosylation of peptide hormones altered the inherent proteolytic instability of this class of biomolecules, the present inventors subjected selected glycopeptide hormones to ex vivo degradation assays using human plasma and in vitro degradation using neprilysin (NEP), insulin degrading enzyme (IDE) and dipeptidyl peptidase IV (DPP-IV) enzymes known to degrade peptide hormones and other bioactive peptides in vivo including ANP, GLP-1, PYY, VIP, Secretin and Galanin.sup.89,90 (
[0212] Degradation Assay:
[0213] For enzymatic degradation assays using either NEP or DPP-IV (R&D systems, UK), an enzyme titration was carried out to ensure full degradation within one hour of reaction time. 15 pM of non-glycosylated peptide substrate and either 50 mM Tris, pH 9, 0.05% Brij for NEP or 50 mM Tris, pH 8 for DPP-IV was treated with varying enzyme amounts in a total volume of 10 μL incubated at 37° C. The degradation assay of the three glycoforms (Tn, T and ST) was performed under same reaction-parameters along with the non-glycosylated peptide (four separate reactions). The following amounts of enzyme were used for NEP reactions: 150 pg/pL enzyme for VIP, Galanin, secretin and their glycoforms, 20 ng/pL for PYY and its glycoforms. 4 and 10 ng/pL DPIV was used for the degradation of VIP+glycoforms and PYY+glycoforms respectively. For ex vivo plasma degradation assays, plasma was diluted to a final concentration of 20% plasma, 50 mM Tris (pH 7.7) and degradation of 15 pM (glyco-) peptide substrate was investigated. Degradation was carried out at 37° C. and several aliquots were taken between 0 minutes and 24 hours and degradation was monitored by MALDI-TOF-MS.
[0214] MALDI-TOF-MS was performed on a Bruker Autoflex instrument (Bruker Daltonik GmbH, Bremen, Germany) by mixing the quenched aliquots with a saturated solution of α-Cyano-4-hydroxycinnamic acid in ACN/H.sub.2O/TFA (70:30:0.1) at a ratio 1:1 on a target steel plate and mass-spectra were acquired in linear mode.
Example 17
O-Glycans on ANP Modulate Stability In Vitro
[0215] For the degradation assays with (glyco-)ANP, the amount of recombinant enzyme used was optimized to fully digest the naked peptide of interest within one hour of incubation at 37° C. In vitro cleavage activity was assayed by adding 2.5 ng Neprilysin (R&D Systems) or 125 ng Insulin-degrading enzyme (IDE, R&D Systems) to 813 pmol peptide or glycopeptide substrate in a total volume of 25 μL. Reactions were performed in 50 mM Tris, 0.05% Brij-35, pH 9 (Neprilysin), or 50 mM Tris, 20 mM NaCl, pH 7.5 (IDE) and incubated at 37° C. Product development was evaluated after 15 min, 30 min, 60 min, and 24 hours by MALDI-TOF and reverse-phase HPLC (C18). Neprilysin degraded ANP completely within 15 min, whereas and ANP-S25/Tn were degraded slower with residual full length ANP-S19/Tn detectable even after 60 min and residual ANP-S25/Tn detectable after 15 min. Residual ANP-S19/Tn, -S25/Tn was detectable after 30 min. After 1 hour, non-modified ANP was completely degraded whereas major degradation products of ANP-S19/Tn and ANP-S19/Tn, -S25/Tn remained detectable even after 24 hours incubation (
Example 18
O-Glycans on Secretin Modulate Stability In Vitro
[0216] In a similar manner, NEP completely degraded Secretin 1-27 to 1-22 within 60 minutes. Also here, glycosylation had a protective effect such that Thr34-Tn remained intact after 60 min and Thr34-T after 120 minutes. Again, sialylated Secretin Thr34-ST appeared resistant to NEP degradation even after 24 hrs (
Example 19
O-Glycans on Galanin Modulate Stability In Vitro
[0217] Nonglycosylated Galanin 1-27 was degraded by NEP within 60 minutes whereas the Ser23-Tn and -T extended glycoforms were degraded after 120 min. The sialylated Galanin-Ser55/ST variant remained intact even after 24 hrs in solution suggesting that sialylation of Galanin is necessary for complete protection from NEP degradation, at least in vitro (
Example 20
O-Glycans on VIP Increase Stability In Vitro
[0218] NEP cleaves VIP sequentially at Asp3/Ala4 then Phe6/Thr7, Lys21/Tyr22 and ultimately at Ala18/Val19 in vitro (
Example 21
O-Glycans on VIP Increase Stability In Vitro
[0219] DPP-IV also degrades VIP both in vivo and in vitro initially cleaving off two N-terminal amino acids Ser2/Asp3, then cleaves at Ala4/Val5 and lastly C-terminally at Tyr22/Leu23. Where the non-glycosylated VIP, Thr7-Tn and Thr7-T glycosylated VIP were degraded equally fast within 60 minutes, the sialylated VIP-Thr7/ST remained fully intact after 30 minutes and partially intact after 60 minutes (
Example 22
O-Glycans on PYY Increase Stability In Vitro
[0220] NEP cleaves PYY at 4 positions at Tyr20/Tyr21, Ser23/Leu24, His26/Tyr27 and Leu30/Val31 in a sequence the present inventors were not able to decide. However whereas non-glycosylated PYY was degraded after 15 minutes all three PYY-Thr32/Tn/T/ST glycosylated peptides remained intact up to 120 minutes (
Example 23
O-Glycans on PYY Increase Stability In Vitro
[0221] DPP-IV cleaves PYY both in vivo and in vitro N-terminally at Pro2/Ile3. Subjecting the non-glycosylated and Thr32 glycopeptide variants to in vitro DPPIV degradation revealed that where Tn- and T-glycosylation had no effect on DPP-IV activity Thr32/ST weakly protected the peptide from N-terminal degradation (
Example 24
O-Glycans on PYY Increase Stability Ex Vivo
[0222] PYY is quickly removed from circulation due to the action of a number of proteases. To approximate in vivo conditions the present inventors chose to analyse PYY degradation using human plasma ex vivo. In plasma PYY is degraded both N-terminally at Pro2/Ile3 and Pro5/Glu6 and C-terminally at Gln34/Arg35 and where O-glycans at residue Thr32 (Thr32/Tn/T/ST) had no effect on the N-terminal degradation, as seen for the in vitro DPP-IV degradation, the present inventors observed full protection from the C-terminal degradation up to 24 hours of the PYY-Thr32/Tn/T/ST glycosylated PYY peptides whereas the non-glycosylated peptide was C-terminally degraded only after 1 hour (
Example 25
O-Glycans on GLP-1.SUB.7-36 .Increase Stability In Vitro and Predicts Increased Stability in Vivo
[0223] DPP-IV is one of the primary enzymes degrading GLP-1.sub.7-36 (from hereon GLP-1) in the circulation in vivo.sup.91. DPP-IV removes the two N-terminal amino acids by cleaving between Ala2/Ser3 both in vitro and in vivo thus inactivating GLP-1, and mutating this DPP-IV cleavage site greatly enhances GLP-1 half-life in vivo.sup.92. Due to this link, DPP-IV inhibitors have successfully been used therapeutically to enhance the effects of endogenous GLP-1.sup.91. To test the effect of glycosylation in close proximity to the DPP-IV cleavage site we incubated GLP-1 with and without glycans with DPP-IV in vitro. Where the non-glycosylated GLP-1 was fully degraded after 30 minutes incubation with equimolar amounts of monoglycosylated GLP-1-Thr5/Tn and GLP-1-Thr7/Tn and equimolar amounts of GLP-1-Thr5/T or GLP-1-Thr7/T were protected against degradation until 120 minutes incubation. Equimolar amounts of the sialylated monoglycosylated GLP-1-Thr5/ST or GLP-1-Thr7/ST remained fully intact until the 120 minute-timepoint and small amounts of intact silaylated GLP-1 was detectable even after 24 h of incubation. The large body of literature on DPP-IV resistant GLP-1 analogs.sup.91-93 taken together with the presented data on Tn, T or ST glycosylated GLP-1, predict that the GLP-1 decorated with either GalNAc (Tn), Gal-GalNAc (T) or Sialyl-GalGalNAc on positions Thr5 or Thr7 has an extended half-life in vivo. Results are summarized in
[0224] NEP also play a role in the degradation of GLP-1 in the circulation, and it has been suggested that NEP is responsible for up to 50% of the degradation of GLP-1.sup.93. In our in vitro assay, NEP cleaves GLP-1 initially at Trp25/Leu26 followed by a second cleavage at Glu21/Phe22 consistent with NEP cleavage sites on GLP-1 reported earlier.sup.94 (
Example 26
O-Glycans on PYY and VIP are Present in Low Stoichiometry in Porcine Intestinal Extracts
[0225] Where the shotgun glycoproteomics strategy is designed to sequence and identify individual O-glycosylation sites it does not allow us to determine site occupancy in a given protein, i.e what is the proportion of glycosylated protein in the total pool of that protein in a given system (e.g. blood, lymph fluid, cell lysate, tissue etc.). To answer this question the present inventors developed a technique to quantify endogenous glycopeptides using either sensitive LC-MS or radioimmunaassay (RIA). Using extracted proteins from pig ileum the present inventors separated proteins from glycoproteins using Jacalin-LWAC and quantified non-glycosylated and glycosylated PYY using either a sensistive PYY-RIA.sup.95 or in the case of VIP, sensitive LC-MS by comparing to isotope labelled standards in the form of in vitro synthesized (glyco-)VIP. In order to prepare the extract for mass spectrometry, the extracted proteins as well as the standards were digested with trypsin prior to Jacalin-LWAC in the case of LC-MS analysis of site occupancy on VIP.
[0226] The present inventors identified approximately 1% glycosylated PYY-Thr32/T and VIP-Thr7-T in porcine ileum tissue ethanol extracts confirming that (sialylated)-T-PYY and -VIP exist in the porcine intestine presumably at a concentration approximately two orders of magnitude below the non-glycosylated.
Example 27
O-Glycans on ANP Confer Retained and Prolonged Differential Agonist Effect In Vivo
[0227] To evaluate the function of O-glycans on ANP in vivo we investigated the cGMP generating, renal and blood pressure actions of equimolar dose (600 pmol/kg/min) ANP-Ser19/ST or ANP-Ser25/ST [n=4/group] in male Sprague Dawley rats (250-350 grams; Charles River Laboratories, Wilmington, Mass.). The protocol is outlined in
[0228] Anesthesia in rats was induced with 133 mg/kg i.p. inactin (Sigma, St Louis, Mo.) and rats were maintained on a heating pad for 1 hour until completely anesthetized. Rats were then subjected to vessel and bladder cannulation for peptide infusion, BP measurement, blood sampling and urine collection. A polyethylene (PE)-5G tube catheter was placed into the jugular vein for inulin, peptide intravenous infusion. The carotid artery was cannulated with a PE-50 tube catheter for BP measurement (Sonometrics, London, Ontario, Canada) and blood sampling. The bladder was accessed and cannulated with a PE-50 tube catheter for passive urine collection. After completion of the above procedural set up, a 45 min equilibration period was performed that included continuous IV inulin and saline infusion. After the 45 min equilibration period, baseline (Time==Q min) parameters were recorded and one blood sampling (0.7 ml) was performed. The inulin and saline infusion was replaced by a continuous intravenously (i.v.) infusion of equimolar ANP-Ser19/ST or ANP-Ser25/ST for 60 min. The infusion rate was weight adjusted and equals weight*0.7/6000 ml/min. After 60 min infusion (Time=60 min), another blood sampling (0.7 ml) was conducted. A post-infusion clearance (Time=90 min) was performed for 30 min. At the end of the study, blood was collected to determine plasma ANP and cGMP levels and to calculate glomerular filtration rate (GFR). Urine was collected at the end of the infusion (Time=60 min) and the study (Time=90 min). Urinary sodium was measured with pHOx Ultra (Nova Biomedical, Waltham, Mass.). Urine flow (UV) and urinary sodium excretion (UNaV) were calculated as urine volume or sodium clearance per min, Inulin concentrations were measured with enthrone method and inulin clearance was used for GFR quantification. Urinary cGMP and ANP excretion rate was calculated based on raw values obtained in the urine and UV.
In Vivo Cardiovascular and Renal Actions
[0229] Infusion with ANP, ANP-Ser19/ST and ANP-Ser25/ST resulted in decrease in mean arterial pressure (MAP) and increase in plasma cGMP over the 60 minutes infusion period (
[0230] Infusion with ANP resulted in an 4-5 fold increase in urine volume (UV) measured after 60 minutes (
Example 28
O-Glycans on ANP Increase Stability In Vivo
[0231] The sustained cardiovascular effect seen with ANP-Ser19/ST and ANP-Ser25/ST suggested that the glycosylated peptides circulate longer compared to the non-glycosylated ANP. Measuring plasma ANP after 90 minutes demonstrated that the non-glycosylated ANP circulated at low concentrations (mean, 148 ng/mL) whereas ANP-Ser19/ST and ANP-Ser25/ST was still present at 17-33 fold higher concentrations (
Comparison of Identified Glycosylated Peptide Hormones to Published Glycosylated Peptide Hormones
[0232] A number of pro-peptide hormones have been reported to be O-glycosylated on the pro-part (non-matured) (pro-brain natriuretic peptide (proBNP), POMC, proglucagon and kininogen). Very recently, two reports describing mature insulin, somatostatin and amylin as well as calcitonin O-glycosylation were released.sup.13,83. Apart from these, only adiponectin has been described as being O-glycosylated before in mammalian studies.
Definition of Peptide Hormones/Neuropeptides/Regulatory Peptides
[0233] Peptide hormones, regulatory peptides or neuropeptides are bioactive peptides that are approximately 3-100 amino acids long. They are involved in cell-cell signaling where they can bind and activate highly specific peptide hormone receptors upon binding.
TABLE-US-00004 TABLE 4 (Human peptide hormones the SEQ ID Nos. of which are disclosed in Tables 6A to 6E infra) ID Sequence Length Family Name NP00002 SVPHFSDEDKDPE 13 7B2 C-terminal peptide (By similarity) NP00012 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQAMNLVGPQSIEGGAHEGL 186 7B2 Neuroendocrine protein 7B2 QHLGPFGNIPNIVAELTGDNIPKDFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAE FSREFQLHQHLFDPEHDYPGLGKWNKKLLYEKMKGGERRKRRSVNPYLQGQRLD NVVAKKSVPHFSDEDKDPE NP00013 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQAMNLVGPQSIEGGAHEGL 150 7B2 N-terminal peptide (By QHLGPFGNIPNIVAELTGDNIPKDFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAE similarity) FSREFQLHQHLFDPEHDYPGLGKWNKKLLYEKMKGGE NP00026 MALQADFDRAAEDVRKLKARPDDGELKELYGLYKQAIVGDINIACPGMLDLKGK 88 ACBP Acyl-CoA-binding domain- AKWEAWNLKKGLSTEDATSAYISKAKELIEKYGI containing protein 7 NP00034 SQAEFEKAAEEVRHLKTKPSDEEMLFIYGHYKQATVGDINTERPGMLDFTGKAKW 86 ACBP Acyl-CoA-binding protein DAWNELKGTSKEDAMKAYINKVEELKKKYGI NP00042 TQAQLLRVGCVLGTCQVQNLSHRLWQLMGPAGRQDSAPVDPSSPHSY 47 Adrenomedullin Adrenomedullin-2 (By similarity) NP00043 VGCVLGTCQVQNLSHRLWQLMGPAGRQDSAPVDPSSPHSY 40 Adrenomedullin Intermedin-short (Potential) NP00044 YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTDKDKDNVAPRSKISPQGY 52 Adrenomedullin Adrenomedullin NP00045 ARLDVASEFRKKWNKWALSR 20 Adrenomedullin Proadrenomedullin N-20 terminal peptide NP00601 QRPRLSHKGPMPF 13 Apelin Apelin-13 (By similarity) NP00602 NGPGPWQGGRRKFRRQRPRLSHKGPMPF 28 Apelin Apelin-28 (By similarity) NP00603 GSRNGPGPWQGGRRKFRRQRPRLSHKGPMPF 31 Apelin Apelin-31 (By similarity) NP00604 LVQPRGSRNGPGPWQGGRRKFRRQRPRLSHKGPMPF 36 Apelin Apelin-36 (By similarity) NP00790 VPLPAGGGTVLTKMYPRGNHWAVGHLM 27 Bombesin/neuromedin- Gastrin-releasing peptide B/ranatensin NP00791 GNHWAVGHLM 10 Bombesin/neuromedin- Neuromedin-C B/ranatensin NP00792 GNLWATGHFM 10 Bombesin/neuromedin- Neuromedin-B B/ranatensin NP00793 APLSWDLPEPRSRASKIRVHSRGNLWATGHFM 32 Bombesin/neuromedin- Neuromedin-B-32 B/ranatensin NP00811 RPPGFSPFR 9 Bradykinin Bradykinin NP00812 KRPPGFSPFR 10 Bradykinin Lysyl-bradykinin NP00813 ISLMKRPPGFSPFR 14 Bradykinin T-kinin NP00846 CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP 32 Calcitonin Calcitonin NP00847 DMSSDLERDHRPHVSMPQNAN 21 Calcitonin Katacalcin NP00848 ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSKAF 37 Calcitonin Calcitonin gene-related peptide 1 NP00849 ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSKAF 37 Calcitonin Calcitonin gene-related peptide 2 NP00850 KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY 37 Calcitonin Islet amyloid polypeptide NP00869 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKKLKS 39 CART CART(1-39) NP00870 VPIYEKKYGQVPMCDAGEQCAVRKGARIGKLCDCPRGTSCNSFLLKCL 48 CART CART(42-89) NP00881 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKKLKSKRVPIYEKKYGQVPMCD 89 CART Cocaine- and amphetamine- AGEQCAVRKGARIGKLCDCPRGTSCNSFLLKCL regulated NP00904 QNDTEPIVLEGKCLVVCDSNPATDSKGSSSSPLGISVRAANSKVAFSAVRSTNHEPS 174 Cerebellins Cerebellin-4 EMSNKTRIIYFDQILVNVGNFFTLESVFVAPRKGIYSFSFHVIKVYQSQTIQVNLMLN GKPVISAFAGDKDVTREAATNGVLLYLDKEDKVYLKLEKGNLVGGWQYSTFSGFL VFPL NP00909 GSAKVAFSAIRSTNH 15 Cerebellins [des-Ser1]-cerebellin NP00910 SGSAKVAFSAIRSTNH 16 Cerebellins Cerebellin NP00911 QNETEPIVLEGKCLVVCDSNPTSDPTGTALGISVRSGSAKVAFSAIRSTNHEPSEMSN 172 Cerebellins Cerebellin-1 RTMIIYFDQVLVNIGNNFDSERSTFIAPRKGIYSFNFHVVKVYNRQTIQVSLMLNGW PVISAFAGDQDVTREAASNGVLIQMEKGDRAYLKLERGNLMGGWKYSTFSGFLVF PL NP00912 QNDTEPIVLEGKCLVVCDSSPSADGAVTSSLGISVRSGSAKVAFSATRSTNHEPSEM 173 Cerebellins Cerebellin-2 SNRTMTIYFDQVLVNIGNHFDLASSIFVAPRKGIYSFSFHVVKVYNRQTIQVSLMQN GYPVISAFAGDQDVTREAASNGVLLLMEREDKVHLKLERGNLMGGWKYSTFSGF LVFPL NP00913 QEGSEPVLLEGECLVVCEPGRAAAGGPGGAALGEAPPGRVAFAAVRSHHHEPAGE 173 Cerebellins Cerebellin-3 TGNGTSGAIYFDQVLVNEGGGFDRASGSFVAPVRGVYSFRFHVVKVYNRQTVQVS LMLNTWPVISAFANDPDVTREAATSSVLLPLDPGDRVSLRLRRGNLLGGWKYSSFS GFLIFPL NP00983 AYGFRGPGPQL 11 Chromogranin/ AL-11 secretogranin NP00984 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQECFETLRGDERILSILRHQNLL 439 Chromogranin/ Chromogranin-A KELQDLALQGAKERAHQQKKHSGFEDELSEVLENQSSQAELKEAVEEPSSKDVME secretogranin KREDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQAPGEEEEEEEEATNTHPPA SLPSQKYPGPQAEGDSEGLSQGLVDREKGLSAEPGWQAKREEEEEEEEEAEAGEEA VPEEEGPTVVLNPHPSLGYKEIRKGESRSEALAVDGAGKPGAEEAQDPEGKGEQEH SQQKEEEEEMAVVPQGLFRGGKSGELEQEEERLSKEWEDSKRWSKMDQLAKELT AEKRLEGQEEEEDNRDSSMKLSFRARAYGFRGPGPQLRRGWRPSSREDSLEAGLPL QVRGYPEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRRG NP00985 EDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQAPGEEEEEEEEATNTHPPASL 92 Chromogranin/ EA-92 PSQKYPGPQAEGDSEGLSQGLVDREKGLSAEPGWQA secretogranin NP00986 EEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRR 37 Chromogranin/ ER-37 secretogranin NP00987 EEEEEEEEEAEAGEEAVPEEEGPTVVLNPHPSL 33 Chromogranin/ ES-43 secretogranin NP00988 GYPEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRR 44 Chromogranin/ GR-44 secretogranin NP00989 GWRPSSREDSLEAGLPLQV 19 Chromogranin/ GV-19 secretogranin NP00990 LEGQEEEEDNRDSSMKLSF 19 Chromogranin/ LF-19 secretogranin NP00991 SEALAVDGAGKPGAEEAQDPEGKGEQEHSQQKEEEEEMAVVPQGLFRG 48 Chromogranin/ Pancreastatin secretogranin NP00992 SGELEQEEERLSKEWEDS 18 Chromogranin/ SS-18 secretogranin NP00993 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQECFETLRGDERILSILRHQNLL 76 Chromogranin/ Vasostatin-1 KELQDLALQGAKERAHQQ secretogranin NP00994 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQECFETLRGDERILSILRHQNLL 113 Chromogranin/ Vasostatin-2 KELQDLALQGAKERAHQQKKHSGFEDELSEVLENQSSQAELKEAVEEPSSKDVME secretogranin NP00995 WSKMDQLA 8 Chromogranin/ WA-8 secretogranin NP00996 WSKMDQLAKELTAE 14 Chromogranin/ WE-14 secretogranin NP00997 SAEFPDFYDSEEPVSTHQEAENEKDRADQTVLTEDEKKELENLAAMDLELQKIAEK 57 Chromogranin/ CCB peptide F secretogranin NP00998 FLGEGHHRVQENQMDKARRHPQGAWKELDRNYLNYGEEGAPGKWQQQGDLQD 74 Chromogranin/ GAWK peptide TKENREEARFQDKQYSSHHTAE secretogranin NP00999 MPVDNRNHNEGMVTRCIIEVLSNALSKSSAPPITPECRQVLKTSRKDVKDKETTEN 657 Chromogranin/ Secretogranin-1 ENTKFEVRLLRDPADASEAHESSSRGEAGAPGEEDIQGPTKADTEKWAEGGGHSRE secretogranin RADEPQWSLYPSDSQVSEEVKTRHSEKSQREDEEEEEGENYQKGERGEDSSEEKHL EEPGETQNAFLNERKQASAIKKEELVARSETHAAGHSQEKTHSREKSSQESGEETG SQENHPQESKGQPRSQEESEEGEEDATSEVDKRRTRPRHHHGRSRPDRSSQGGSLPS EEKGHPQEESEESNVSMASLGEKRDHHSTHYRASEEEPEYGEEIKGYPGVQAPEDL EWERYRGRGSEEYRAPRPQSEESWDEEDKRNYPSLELDKMAHGYGEESEEERGLE PGKGRHHRGRGGEPRAYFMSDTREEKRFLGEGHHRVQENQMDKARRHPQGAWK ELDRNYLNYGEEGAPGKWQQQGDLQDTKENREEARFQDKQYSSHHTAEKRKRLG ELFNPYYDPLQWKSSHFERRDNMNDNFLEGEEENELTLNEKNFFPEYNYDWWEK KPFSEDVNWGYEKRNLARVPKLDLKRQYDRVAQLDQLLHYRKKSAEFPDFYDSE EPVSTHQEAENEKDRADQTVLTEDEKKELENLAAMDLELQKIAEKFSQRG NP01000 QRNQLLQKEPDLRLENVQKFPSPEMIRALEYIENLRQQAHKEESSPDYNPYQGVSV 587 Chromogranin/ Secretogranin-2 PLQQKENGDESHLPERDSLSEEDWMRIILEALRQAENEPQSAPKENKPYALNSEKN secretogranin FPMDMSDDYETQQWPERKLKHMQFPPMYEENSRDNPFKRTNEIVEEQYTPQSLAT LESVFQELGKLTGPNNQKRERMDEEQKLYTDDEDDIYKANNIAYEDVVGGEDWN PVEEKIESQTQEEVRDSKENIEKNEQINDEMKRSGQLGIQEEDLRKESKDQLSDDVS KVIAYLKRLVNAAGSGRLQNGQNGERATRLFEKPLDSQSIYQLIEISRNLQIPPEDLI EMLKTGEKPNGSVEPERELDLPVDLDDISEADLDHPDLFQNRMLSKSGYPKTPGRA GTEALPDGLSVEDILNLLGMESAANQKTSYFPNPYNQEKVLPRLPYGAGRSRSNQL PKAAWIPHVENRQMAYENLNDKDQELGEYLARMLVKYPEIINSNQVKRVPGQGSS EDDLQEEEQIEQAIKEHLNQGSSQETDKLAPVSKRFPVGPPKNDDTPNRQYWDEDL LMKVLEYLNQEKAEKGREHIAKRAMENM NP01001 TNEIVEEQYTPQSLATLESVFQELGKLTGPNNQ 33 Chromogranin/ Secretoneurin secretogranin NP01002 FPKPGGSQDKSLHNRELSAERPLNEQIAEAEEDKIKKTYPPENKPGQSNYSFVDNLN 449 Chromogranin/ Secretogranin-3 LLKAITEKEKIEKERQSIRSSPLDNKLNVEDVDSTKNRKLIDDYDSTKSGLDHKFQD secretogranin DPDGLHQLDGTPLTAEDIVHKIAARIYEENDRAVFDKIVSKLLNLGLITESQAHTLE DEVAEVLQKLISKEANNYEEDPNKPTSWTENQAGKIPEKVTPMAAIQDGLAKGEN DETVSNTLTLTNGLERRTKTYSEDNFEELQYFPNFYALLKSIDSEKEAKEKETLITIM KTLIDFVKMMVKYGTISPEEGVSYLENLDEMIALQTKNKLEKNATDNISKLFPAPSE KSHEETDSTKEEAAKMEKEYGSLKDSTKDDNSNPGGKTDEPKGKTEAYLEAIRKNI EWLKKHDKKGNKEDYDLSKMRDFINKQADAYVEKGILDKEEAEAIKRIYSSL NP01097 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFVLYRITEATKTVGSDTFYSF 626 Cystatin Kininogen-1 KYEIKEGDCPVQSGKTWQDCEYKDAAKAATGECTATVGKRSSTKFSVATQTCQIT PAEGPVVTAQYDCLGCVHPISTQSPDLEPILRHGIQYFNNNTQHSSLFMLNEVKRAQ RQVVAGLNFRITYSIVQTNCSKENFLFLTPDCKSLWNGDTGECTDNAYIDIQLRIAS FSQNCDIYPGKDFVQPPTKICVGCPRDIPTNSPELEETLTHTITKLNAENNATFYFKID NVKKARVQVVAGKKYFIDFVARETTCSKESNEELTESCETKKLGQSLDCNAEVYV VPWEKKIYPTVNCQPLGMISLMKRPPGFSPFRSSRIGEIKEETTVSPPHTSMAPAQDE ERDSGKEQGHTRRHDWGHEKQRKHNLGHGHKHERDQGHGHQRGHGLGHGHEQ QHGLGHGHKFKLDDDLEHQGGHVLDHGHKHKHGHGHGKHKNKGKKNGKHNG WKTEHLASSSEDSTTPSAQTQEKTEGPTPIPSLAKPGVTVTFSDFQDSDLIATMMPPI SPAPIQSDDDWIPDIQIDPNGLSFNPISDFPDTTSPKCPGRPWKSVSEINPTTQMKESY YFDLTDGLS NP01098 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFVLYRITEATKTVGSDTFYSF 362 Cystatin Kininogen-1 heavy chain KYEIKEGDCPVQSGKTWQDCEYKDAAKAATGECTATVGKRSSTKFSVATQTCQIT PAEGPVVTAQYDCLGCVHPISTQSPDLEPILRHGIQYFNNNTQHSSLFMLNEVKRAQ RQVVAGLNFRITYSIVQTNCSKENFLFLTPDCKSLWNGDTGECTDNAYIDIQLRIAS FSQNCDIYPGKDFVQPPTKICVGCPRDIPTNSPELEETLTHTITKLNAENNATFYFKID NVKKARVQVVAGKKYFIDFVARETTCSKESNEELTESCETKKLGQSLDCNAEVYV VPWEKKIYPTVNCQPLGMISLMK NP01099 SSRIGEIKEETTVSPPHTSMAPAQDEERDSGKEQGHTRRHDWGHEKQRKHNLGHG 255 Cystatin Kininogen-1 light chain HKHERDQGHGHQRGHGLGHGHEQQHGLGHGHKFKLDDDLEHQGGHVLDHGHK HKHGHGHGKHKNKGKKNGKHNGWKTEHLASSSEDSTTPSAQTQEKTEGPTPIPSL AKPGVTVTFSDFQDSDLIATMMPPISPAPIQSDDDWIPDIQIDPNGLSFNPISDFPDTT SPKCPGRPWKSVSEINPTTQMKESYYFDLTDGLS NP01100 WGHE 4 Cystatin Low molecular weight growth-promoting factor NP01125 CSCSSLMDKECVYFCHLDIIWVNTPEHVVPYGLGSPRS 38 Endothelin/sarafotoxin Big endothelin-1 NP01126 CSCSSLMDKECVYFCHLDIIW 21 Endothelin/sarafotoxin Endothelin-1 NP01127 CSCSSWLDKECVYFCHLDIIW 21 Endothelin/sarafotoxin Endothelin-2 NP01128 CTCFTYKDKECVYYCHLDIIW 21 Endothelin/sarafotoxin Endothelin-3 NP01760 AGEGLNSQFWSLAAPQRF 18 FMRFamide related Neuropeptide AF peptide NP01761 FLFQPQRF 8 FMRFamide related Neuropeptide FF peptide NP01762 SQAFLFQPQRF 11 FMRFamide related Neuropeptide SF peptide NP01763 SLNFEELKDWGPKNVIKMSTPAVNKMPHSFANLPLRF 37 FMRFamide related Neuropeptide NPSF peptide (Potential) NP01764 VPNLPQRF 8 FMRFamide related Neuropeptide NPVF peptide NP01765 MPHSFANLPLRF 12 FMRFamide related Neuropeptide RFRP-1 peptide NP01766 SAGATANLPLRS 12 FMRFamide related Neuropeptide RFRP-2 peptide (Potential) NP02007 TPDINPAWYASRGIRPVGRF 20 FMRFamide related Prolactin-releasing peptide peptide PrRP20 NP02008 SRTHRHSMEIRTPDINPAWYASRGIRPVGRF 31 FMRFamide related Prolactin-releasing peptide peptide PrRP31 NP02025 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 30 Galanin Galanin NP02026 ELRPEDDMKPGSFDRSIPENNIMRTIIEFLSFLHLKEAGALDRLLDLPAAASSEDIERS 59 Galanin Galanin message-associated peptide NP02040 APAHRGRGGWTLNSAGYLLGPVLHLPQMGDQDGKRETALEILDLWKAIDGLPYS 60 Galanin Galanin-like peptide HPPQPS NP02242 QPVPPADPAGSGLQRAEEAPRRQLRVSQRTDGESRAHLGALLARYIQQARKAPSGR 95 Gastrin/cholecystokinin Cholecystokinin MSIVKNLQNLDPSHRISDRDYMGWMDFGRRSAEEYEYPS NP02243 ISDRDYMGWMDF 12 Gastrin/cholecystokinin Cholecystokinin-12 NP02244 LDPSHRISDRDYMGWMDF 18 Gastrin/cholecystokinin Cholecystokinin-18 (By similarity) NP02245 IVKNLQNLDPSHRISDRDYMGWMDF 25 Gastrin/cholecystokinin Cholecystokinin-25 (By similarity) NP02246 KAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 33 Gastrin/cholecystokinin Cholecystokinin-33 NP02247 YIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 39 Gastrin/cholecystokinin Cholecystokinin-39 NP02248 GWMDF 5 Gastrin/cholecystokinin Cholecystokinin-5 (By similarity) NP02249 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYMGW 58 Gastrin/cholecystokinin Cholecystokinin-58 MDF NP02250 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKNLQNLDPSHRISD 49 Gastrin/cholecystokinin Cholecystokinin-58 desnonopeptide (By similarity) NP02251 YMGWMDF 7 Gastrin/cholecystokinin Cholecystokinin-7 (By similarity) NP02252 DYMGWMDF 8 Gastrin/cholecystokinin Cholecystokinin-8 NP02253 QLGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF 34 Gastrin/cholecystokinin Big gastrin NP02254 QGPWLEEEEEAYGWMDF 17 Gastrin/cholecystokinin Gastrin NP02255 WLEEEEEAYGWMDF 14 Gastrin/cholecystokinin Gastrin-14 NP02256 DLELPWLEQQGPASHHRRQLGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF 52 Gastrin/cholecystokinin Gastrin-52 NP02257 YGWMDF 6 Gastrin/cholecystokinin Gastrin-6 NP02258 SWKPRSQQPDAPLGTGANRDLELPWLEQQGPASHHRRQLGPQGPPHLVADPSKKQ 71 Gastrin/cholecystokinin Gastrin-71 GPWLEEEEEAYGWMDF NP02464 YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ 42 Glucagon Gastric inhibitory polypeptide NP02465 RSLQDTEEKSRSFSASQADPLSDPDQMNEDKRHSQGTFTSDYSKYLDSRRAQDFVQ 69 Glucagon Glicentin (By similarity) WLMNTKRNRNNIA NP02466 RSLQDTEEKSRSFSASQADPLSDPDQMNED 30 Glucagon Glicentin-related polypeptide (By similarity) NP02467 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 29 Glucagon Glucagon NP02468 HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 37 Glucagon Glucagon-like peptide 1 NP02469 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 30 Glucagon Glucagon-like peptide 1(7-36) NP02470 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 31 Glucagon Glucagon-like peptide 1(7-37) NP02471 HADGSFSDEMNTILDNLAARDFINWLIQTKITD 33 Glucagon Glucagon-like peptide 2 (By similarity) NP02472 HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA 37 Glucagon Oxyntomodulin (By similarity) NP02473 DVAHGILNEAYRKVLDQLSAGKHLQSLVARGVGGSLGGGAGDDAEPLS 48 Glucagon PACAP-related peptide NP02474 HSDGIFTDSYSRYRKQMAVKKYLAAVL 27 Glucagon Pituitary adenylate cyclase- activating polypeptide 27 NP02475 HSDGIFTDSYSRYRKQMAVKKYLAAVLGKRYKQRVKNK 38 Glucagon Pituitary adenylate cyclase- activating polypeptide 38 NP02476 HSDGTFTSELSRLREGARLQRLLQGLV 27 Glucagon Secretin NP02477 YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL 44 Glucagon Somatoliberin NP02478 HADGVFTSDFSKLLGQLSAKKYLESLM 27 Glucagon Intestinal peptide PHM-27 NP02479 HADGVFTSDFSKLLGQLSAKKYLESLMGKRVSSNISEDPVPV 42 Glucagon Intestinal peptide PHV-42 NP02480 HSDAVFTDNYTRLRKQMAVKKYLNSILN 28 Glucagon Vasoactive intestinal peptide NP02572 DAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLRDLKGALESLIEEETGQKKI 56 GnRH GnRH-associated peptide 1 NP02573 QHWSYGLRPG 10 GnRH Gonadoliberin-1 NP02574 QHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLRDLKGAL 69 GnRH Progonadoliberin-1 ESLIEEETGQKKI NP02575 ALSSAQDPQNALRPPGRALDTAAGSPVQTAHGLPSDALAPLDDSMPWEGRTTAQ 84 GnRH GnRH-associated peptide 2 WSLHRKRHLARTLLTAAREPRPAPPSSNKV NP02576 QHWSHGWYPG 10 GnRH Gonadoliberin-2 NP02577 QHWSHGWYPGGKRALSSAQDPQNALRPPGRALDTAAGSPVQTAHGLPSDALAPL 97 GnRH Progonadoliberin-2 DDSMPWEGRTTAQWSLHRKRHLARTLLTAAREPRPAPPSSNKV NP02765 GPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSRRAPQTGIVDECCFRSCDLRR 70 Insulin Insulin-like growth factor I LEMYCAPLKPAKSA NP02766 AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRGIVEECCFRSCDLALLE 67 Insulin Insulin-like growth factor II TYCATPAKSE NP02767 YRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRGIVEECCFRSCDLALLET 66 Insulin Insulin-like growth factor II YCATPAKSE Ala-25 Del NP02768 DVSTPPTVLPDNFPRYPVGKFFQYDTWKQSTQRL 34 Insulin Preptin NP02769 GIVEQCCTSICSLYQLENYCN 21 Insulin Insulin A chain NP02770 FVNQHLCGSHLVEALYLVCGERGFFYTPKT 30 Insulin Insulin B chain NP02771 PYVALFEKCCLIGCTKRSLAKYC 23 Insulin Relaxin A chain (By similarity) NP02772 VAAKWKDDVIKLCGRELVRAQIAICGMSTWS 31 Insulin Relaxin B chain (By similarity) NP02773 QLYSALANKCCHVGCTKRSLARFC 24 Insulin Relaxin A chain NP02774 DSWMEEVIKLCGRELVRAQIAICGMSTWS 29 Insulin Relaxin B chain NP02775 DVLAGLSSSCCKWGCSKSEISSLC 24 Insulin Relaxin-3 A chain (By similarity) NP02776 RAAPYGVRLCGREFIRAVIFTCGGSRW 27 Insulin Relaxin-3 B chain (By similarity) NP02840 YNWNSFGLRF 10 KISS1 Kisspeptin-10 NP02841 LPNYNWNSFGLRF 13 KISS1 Kisspeptin-13 NP02842 DLPNYNWNSFGLRF 14 KISS1 Kisspeptin-14 NP02843 EPLEKVASVGNSRPTGQQLESLGLLAPGEQSLPCTERKPAATARLSRRGTSLSPPPES 119 KISS1 Metastasis-suppressor KiSS-1 SGSPQQPGLSAPHSRQIPAPQGAVLVQREKDLPNYNWNSFGLRFGKREAAPGNHG RSAGRG NP02844 GTSLSPPPESSGSPQQPGLSAPHSRQIPAPQGAVLVQREKDLPNYNWNSFGLRF 54 KISS1 Metastin NP02870 VPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVTGLDFIPGLHPILTLSKMDQTL 146 Leptin Leptin AVYQQILTSMPSRNVIQISNDLENLRDLLHVLAFSKSCHLPWASGLETLDSLGGVLE ASGYSTEVVALSRLQGSLQDMLWQLDLSPGC NP02908 DFDMLRCMLGRVYRPCWQV 19 Melanin-concentrating Melanin-concentrating hormone hormone NP02909 EIGDEENSAKFPI 13 Melanin-concentrating Neuropeptide-glutamic acid- hormone isoleucine NP02910 GSVAFPAENGVQNTESTQE 19 Melanin-concentrating Neuropeptide-glycine- hormone glutamic acid(Potential) NP02937 ILLSASKSIRNLDDDMVFNTFRLGKGFQKEDTAEKSVIAPSLEQYKNDESSFMNEEE 144 Melanin-concentrating Pro-MCH NKVSKNTGSKHNFLNHGLPLNLAIKPYLALKGSVAFPAENGVQNTESTQEKREIGD hormone EENSAKFPIGRRDFDMLRCMLGRVYRPCWQV NP03000 GSSFLSPEHQRVQQRKESKKPPAKLQP 27 Motilin Ghrelins-27 NP03001 GSSFLSPEHQRVQQRKESKKPPAKLQPR 28 Motilin Ghrelins-28 NP03002 FNAPFDVGIKLSGVQYQQHSQAL 23 Motilin Obestatin NP03003 FVPIFTYGELQRMQEKERNKGQ 22 Motilin Motilin NP03004 SLSVWQRSGEEGPVDPAEPIREEENEMIKLTAPLEIGMRMNSRQLEKYPATLEGLLS 66 Motilin Motilin-associated peptide EMLPQHAAK NP03027 FVPIFTYGELQRMQEKERNKGQKKSLSVWQRSGEEGPVDPAEPIREEENEMIKLTA 90 Motilin Promotilin PLEIGMRMNSRQLEKYPATLEGLLSEMLPQHAAK NP03617 ETTTQGPGVLLPLPKGACTGWMAGIPGHPGHNGAPGRDGRDGTPGEKGEKGDPGL 226 NA Adiponectin IGPKGDIGETGVPGAEGPRGFPGIQGRKGEPGEGAYVYRSAFSVGLETYVTIPNMPI RFTKIFYNQQNHYDGSTGKFHCNIPGLYYFAYHITVYMKDVKVSLFKKDKAMLFT YDQYQENNVDQASGSVLLHLEVGDQVWLQVYGEGERNGLYADNDNDSTFTGFLL YHDTN NP03618 MIEVVCNDRLGKKVRVKCNTDDTIGDLKKLIAAQTGTRWNKIVLKKWYTIFKDHV 73 NA Ubiquitin-like protein 5 SLGDYEIHDGMNLELYYQ NP03619 AQMGLAPMEGIRRPDQALLPELPGLGLRAPLKKTTAEQAEEDLLQEAQALAEVLD 112 NA Agouti-related protein LQDREPRSSRRCVRLHESCLGQQVPCCDPCATCYCRFFNAFCYCRKLGTAMNPCSR T NP03624 MNPAAEAEFNILLATDSYKVTHYKQYPPNTSKVYSYFECREKKTENSKLRKVKYE 491 NAPRTase Nicotinamide ETVFYGLQYILNKYLKGKVVTKEKIQEAKDVYKEHFQDDVFNEKGWNYILEKYD phosphoribosyltransferase GHLPIEIKAVPEGFVIPRGNVLFTVENTDPECYWLTNWIETILVQSWYPITVATNSRE QKKILAKYLLETSGNLDGLEYKLHDFGYRGVSSQETAGIGASAHLVNFKGTDTVA GLALIKKYYGTKDPVPGYSVPAAEHSTITAWGKDHEKDAFEHIVTQFSSVPVSVVS DSYDIYNACEKIWGEDLRHLIVSRSTQAPLIIRPDSGNPLDTVLKVLEILGKKFPVTE NSKGYKLLPPYLRVIQGDGVDINTLQEIVEGMKQKMWSIENIAFGSGGGLLQKLTR DLLNCSFKCSYVVTNGLGINVFKDPVADPNKRSKKGRLSLHRTPAGNFVTLEEGKG DLEEYGQDLLHTVFKNGKVTKSYSFDEIRKNAQLNIELEAAHH NP03673 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Natriuretic peptide Atrial natriuretic factor NP03674 NPMYNAVSNADLMDFKNLLDHLEEKMPLED 30 Natriuretic peptide Cardiodilatin-related peptide NP03675 SPKMVQGSGCFGRKMDRISSSSGLGCKV 28 Natriuretic peptide BNP(1-28) NP03676 SPKMVQGSGCFGRKMDRISSSSGLGCKVL 29 Natriuretic peptide BNP(1-29) NP03677 SPKMVQGSGCFGRKMDRISSSSGLGCKVLR 30 Natriuretic peptide BNP(1-30) NP03678 PKMVQGSGCFGRKMDRISSSSGLGCKVLRR 30 Natriuretic peptide BNP(2-31) NP03679 KMVQGSGCFGRKMDRISSSSGLGCKVL 27 Natriuretic peptide BNP(3-29) NP03680 KMVQGSGCFGRKMDRISSSSGLGCKVLR 28 Natriuretic peptide BNP(3-30) NP03681 KMVQGSGCFGRKMDRISSSSGLGCKVLRRH 30 Natriuretic peptide BNP(3-32) NP03682 MVQGSGCFGRKMDRISSSSGLGCK 24 Natriuretic peptide BNP(4-27) NP03683 MVQGSGCFGRKMDRISSSSGLGCKVL 26 Natriuretic peptide BNP(4-29) NP03684 MVQGSGCFGRKMDRISSSSGLGCKVLR 27 Natriuretic peptide BNP(4-30) NP03685 MVQGSGCFGRKMDRISSSSGLGCKVLRR 28 Natriuretic peptide BNP(4-31) NP03686 MVQGSGCFGRKMDRISSSSGLGCKVLRRH 29 Natriuretic peptide BNP(4-32) NP03687 VQGSGCFGRKMDRISSSSGLGCKVL 25 Natriuretic peptide BNP(5-29) NP03688 VQGSGCFGRKMDRISSSSGLGCKVLRR 27 Natriuretic peptide BNP(5-31) NP03689 VQGSGCFGRKMDRISSSSGLGCKVLRRH 28 Natriuretic peptide BNP(5-32) NP03690 SPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH 32 Natriuretic peptide Brain natriuretic peptide 32 NP03691 HPLGSPGSASDLETSGLQEQRNHLQGKLSELQVEQTSLEPLQESPRPTGVWKSREV 108 Natriuretic peptide Natriuretic peptides B ATEGIRGHRKMVLYTLRAPRSPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH NP03692 GLSKGCFGLKLDRIGSMSGLGC 22 Natriuretic peptide CNP-22 NP03693 YKGANKKGLSKGCFGLKLDRIGSMSGLGC 29 Natriuretic peptide CNP-29 NP03694 DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC 53 Natriuretic peptide CNP-53 NP03714 ANLTNGGKSELLKSGSSKSTLKHIWTESSKDLSISRLLSQTFRGKENDTDLDLRYDT 250 Neurexophilin Neurexophilin-1 PEPYSEQDLWDWLRNSTDLQEPRPRAKRRPIVKTGKFKKMFGWGDFHSNIKTVKL NLLITGKIVDHGNGTFSVYFRHNSTGQGNVSVSLVPPTKIVEFDLAQQTVIDAKDSK SFNCRIEYEKVDKATKNTLCNYDPSKTCYQEQTQSHVSWLCSKPFKVICIYISFYST DYKLVQKVCPDYNYHSDTPYFPSG NP03715 KEVVHATEGLDWEDKDAPGTLVGNVVHSRIISPLRLFVKQSPVPKPGPMAYADSM 242 Neurexophilin Neurexophilin-2 ENFWDWLANITEIQEPLARTKRRPIVKTGKFKKMFGWGDFHSNIKTVKLNLLITGKI VDHGNGTFSVYFRHNSTGLGNVSVSLVPPSKVVEFEVSPQSTLETKESKSFNCRIEY EKTDRAKKTALCNFDPSKICYQEQTQSHVSWLCSKPFKVICIYIAFYSVDYKLVQK VCPDYNYHSETPYLSSG NP03716 QDDGPPGSEDPERDDHEGQPRPRVPRKRGHISPKSRPMANSTLLGLLAPPGEAWGI 230 Neurexophilin Neurexophilin-3 LGQPPNRPNHSPPPSAKVKKIFGWGDFYSNIKTVALNLLVTGKIVDHGNGTFSVHF QHNATGQGNISISLVPPSKAVEFHQEQQIFIEAKASKIFNCRMEWEKVERGRRTSLC THDPAKICSRDHAQSSATWSCSQPFKVVCVYIAFYSTDYRLVQKVCPDYNYHSDTP YYPSG NP03717 QIPESGRPQYLGLRPAAAGAGAPGQQLPEPRSSDGLGVGRAWSWAWPTNHTGAL 285 Neurexophilin Neurexophilin-4 ARAGAAGALPAQRTKRKPSIKAARAKKIFGWGDFYFRVHTLKFSLLVTGKIVDHV NGTFSVYFRHNSSSLGNLSVSIVPPSKRVEFGGVWLPGPVPHPLQSTLALEGVLPGL GPPLGMAAAAAGPGLGGSLGGALAGPLGGALGVPGAKESRAFNCHVEYEKTNRA RKHRPCLYDPSQVCFTEHTQSQAAWLCAKPFKVICIFVSFLSFDYKLVQKVCPDYN FQSEHPYFG NP03732 ILQRGSGTAAVDFTKKDHTATWGRPFFLFRPRN 33 Neuromedins Neuromedin-S NP03733 FRVDEEFQSPFASQSRGYFLFRPRN 25 Neuromedins Neuromedin-U-25 NP03744 WYKPAAGHSSYSVGRAAGLLSGLR 24 Neuropeptide B/W Neuropeptide B-23 NP03745 WYKPAAGHSSYSVGRAAGLLSGLRRSPYA 29 Neuropeptide B/W Neuropeptide B-29 NP03746 WYKHVASPRYHTVGRAAGLLMGL 23 Neuropeptide B/W Neuropeptide W-23 NP03747 WYKHVASPRYHTVGRAAGLLMGLRRSPYLW 30 Neuropeptide B/W Neuropeptide W-30 NP03755 SFRNGVGTGMKKTSFQRAKS 20 Neuropeptide S Neuropeptide S NP03782 SDSEEEMKALEADFLTNMHTSKISKAHVPSWKMTLLNVCSLVNNLNSPAEETGEV 125 Neurotensin Large neuromedin N HEEELVARRKLPTALDGFSLEAMLTIYQLHKICHSRAFQHWELIQEDILDTGNDKN GKEEVIKRKIPYIL NP03783 IPYIL 5 Neurotensin Neuromedin N NP03784 QLYENKPRRPYIL 13 Neurotensin Neurotensin NP03785 DSYYY 5 Neurotensin Tail peptide (Potential) NP03886 SSPETLISDLLMRESTENVPRTRLEDPAMW 30 NPY C-flanking peptide of NPY NP03887 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 36 NPY Neuropeptide Y NP03957 APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY 36 NPY Pancreatic hormone NP03958 HKEDTLAFSEWGSPHAAVPR 20 NPY Pancreatic icosapeptide NP03959 YPIKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 36 NPY Peptide YY NP03960 IKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 34 NPY Peptide YY(3-36) NP04021 VPLERGAPNKEETPATESPDTGLYYHRYLQEVIDVLETDGHFREKLQAANAEDIKS 435 Nucleobindin Nucleobindin-1 GKLSRELDFVSHHVRTKLDELKRQEVSRLRMLLKAKMDAEQDPNVQVDHLNLLK QFEHLDPQNQHTFEARDLELLIQTATRDLAQYDAAHHEEFKRYEMLKEHERRRYL ESLGEEQRKEAERKLEEQQRRHREHPKVNVPGSQAQLKEVWEELDGLDPNRFNPK TFFILHDINSDGVLDEQELEALFTKELEKVYDPKNEEDDMREMEEERLRMREHVM KNVDTNQDRLVTLEEFLASTQRKEFGDTGEGWETVEMHPAYTEEELRRFEEELAA REAELNAKAQRLSQETEALGRSQGRLEAQKRELQQAVLHMEQRKQQQQQQQGH KAPAAHPEGQLKFHPDTDDVPVPAPAGDQKEVDTSEKKLLERLPEVEVPQHL NP04025 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVIDVLETDKHFREKLQKADIE 82 Nucleobindin Nesfatin-1 EIKSGRLSKELDLVSHHVRTKLDEL NP04026 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVIDVLETDKHFREKLQKADIE 396 Nucleobindin Nucleobindin-2 EIKSGRLSKELDLVSHHVRTKLDELKRQEVGRLRMLIKAKLDSLQDIGMDHQALLK QFDHLNHLNPDKFESTDLDMLIKAATSDLEHYDKTRHEEFKKYEMMKEHERREYL KTLNEEKRKEEESKFEEMKKKHENHPKVNHPGSKDQLKEVWEETDGLDPNDFDP KTFFKLHDVNSDGFLDEQELEALFTKELEKVYDPKNEEDDMVEMEEERLRMREHV MSEVDTNKDRLVTLEEFLKATEKKEFLEPDSWETLDQQQFFTEEELKEYENIIALQE NELKKKADELQKQKEELQRQHDQLEAQKLEYHQVIQQMEQKKLQQGIPPSGPAGE LKFEPHI NP04038 YAFDVVG 7 Opioid Deltorphin I NP04039 ELTGQRLRQGDGPNAGADDGPGAQADLEHSLLVAAEKKD 57 Opioid Gamma-Lipotropin EGPYRMEHFRWGSPPKD NP04040 YVMGHFRWDRFG 12 Opioid □pmelanocyte-stimulating hormone NP04123 YGGFLRKYPK 10 Opioid Alpha-neoendorphin NP04124 YGGFLRKYP 9 Opioid Beta-neoendorphin NP04125 YGGFLRRIRPKLKWDNQKRYGGFLRRQFKVVT 32 Opioid Big dynorphin NP04126 YGGFLRRIRPKLK 13 Opioid Dynorphin A(1-13) (By similarity) NP04127 YGGFLRRIRPKLKWDNQ 17 Opioid Dynorphin A(1-17) NP04128 YGGFLRRI 8 Opioid Dynorphin A(1-8) (By similarity) NP04129 YGGFL 5 Opioid Leu-enkephalin NP04130 YGGFLRRQFKVVTRSQEDPNAYSGELFDA 29 Opioid Leumorphin NP04131 YGGFLRRQFKVVT 13 Opioid Rimorphin NP04132 YGGFM 5 Opioid Met-enkephalin NP04133 YGGFMRGL 8 Opioid Met-enkephalin-Arg-Gly-Leu NP04134 YGGFMRF 7 Opioid Met-enkephalin-Arg-Phe NP04135 MDELYPMEPEEEANGSEILA 20 Opioid PENK(114-133) (By similarity) NP04136 DAEEDDSLANSSDLLKELLETGDNRERSHHQDGSDNEEEVS 41 Opioid PENK(143-183) (By similarity) NP04137 FAEALPSDEEGESYSKEVPEME 22 Opioid PENK(237-258) (By similarity) NP04138 ECSQDCATCSYRLVRPADINFLACVMECEGKLPSLKIWETCKELLQLSKPELPQDG 73 Opioid Synenkephalin TSTLRENSKPEESHLLA NP04139 MPRVRSLFQEQEEPEPGMEEAGEMEQKQLQ 30 Opioid Neuropeptide 1 (Probable) NP04140 FSEFMRQYLVLSMQSSQ 17 Opioid Neuropeptide 2 (Probable) NP04141 FGGFTGARKSARKLANQ 17 Opioid Nociceptin NP04400 QPLPDCCRQKTCSCRLYELLHGAGNHAAGILTL 33 Orexin Orexin-A NP04401 RSGPPGLQGRLQRLLQASGNHAAGILTM 28 Orexin Orexin-B NP04409 SLALADDAAFRERARLLAALERRHWLNSYMHKLLVLDAP 39 Parathyroid hormone Tuberoinfundibular peptide of 39residues NP04423 TRSAWLDSGVTGSGLEGDHLSDTSTTSLELDSR 33 Parathyroid hormone Osteostatin NP04424 AVSEHQLLHDKGKSIQDLRRRFFLHHLIAEIHTAEIRATSEVSPNSKPSPNTKNHPVR 141 Parathyroid hormone Parathyroid hormone-related FGSDDEGRYLTQETNKVETYKEQPLKTPGKKKKGKPGKRKEQEKKKRRTRSAWL protein DSGVTGSGLEGDHLSDTSTTSLELDSRRH NP04425 AVSEHQLLHDKGKSIQDLRRRFFLHHLIAEIHTAEI 36 Parathyroid hormone PTHrP[1-36] NP04426 ATSEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKVETYKEQPLKTPGKKKKGK 57 Parathyroid hormone PTHrP[38-94] P NP04892 YGGFMTSEKSQTPLVTLFKNAIIKNAYKKGE 31 POMC Beta-endorphin NP04893 SYSMEHFRWGKPVGKKRRPVKVYPNGAEDESAEAFPLEF 39 POMC Corticotropin NP04894 PVKVYPNGAEDESAEAFPLEF 21 POMC Corticotropin-like intermediary peptide NP04895 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAAEKKDEGPYRMEHFRWGSPP 89 POMC Lipotropin beta KDKRYGGFMTSEKSQTPLVTLFKNAIIKNAYKKGE NP04896 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAAEKKDEGPYRMEHFRWGSPP 56 POMC Lipotropin gamma KD NP04897 SYSMEHFRWGKPV 13 POMC Melanotropin alpha NP04898 DEGPYRMEHFRWGSPPKD 18 POMC Melanotropin beta NP04899 YVMGHFRWDRF 11 POMC Melanotropin gamma NP04900 WCLESSQCQDLTTESNLLECIRACKPDLSAETPMFPGNGDEQPLTENPRKYVMGHF 76 POMC NPP RWDRFGRRNSSSSGSSGAGQ NP04901 EDVSAGEDCGPLPEGGPEPRSDGAKPGPRE 30 POMC Potential peptide NP04921 ARPVKEPRGLSAASPPLAETGAPRRFRRSVPRGEAAGAVQELARALAHLLEAERQE 227 ProSAAS ProSAAS RARAEAQEAEDQQARVLAQLLRVWGAPRNSDPALGLDDDPDAPAAQLARALLRA RLDPAALAAQLVPAPVPAAALRPRPPVYDDGPAGPDAEEAGDETPDVDPELLRYL LGRILAGSADSEGVAAPRRLRRAADHDVGSELPPEGVLGALLRVKRLETPAPQVPA RRLLPP NP04922 LETPAPQVPARRLLPP 16 ProSAAS Big LEN (By similarity) NP04923 AADHDVGSELPPEGVLGALLRVKRLETPAPQVPARRLLPP 40 ProSAAS Big PEN-LEN (By similarity) NP04924 ARPVKEPRGLSAASPPLAETGAPRRF 26 ProSAAS Big SAAS (By similarity) NP04925 ARPVKEP 7 ProSAAS KEP (By similarity) NP04926 LETPAPQVPA 10 ProSAAS Little LEN (By similarity) NP04927 GLSAASPPLAETGAPRRF 18 ProSAAS Little SAAS (By similarity) NP04928 AADHDVGSELPPEGVLGALLRV 22 ProSAAS PEN (By similarity) NP05218 KTLCSMEEAINERIQEVAGSLIFRAISSIGLECQSVTSRGDLATCPRGFAVTGCTCGS 90 Resistin/FIZZ Resistin ACGSWDVRAETTCHCQCAGMDWTGARCCRVQP NP05219 QCSLDSVMDKKIKDVLNSLEYSPSPISKKLSCASVKSQGRPSSCPAGMAVTGCACG 88 Resistin/FIZZ Resistin-like beta YGCGSWDVQLETTCHCQCSVVDWTTARCCHLT NP05222 QDEGSEATGFLPAAGEKTSGPLGNLAEELNGYSRKKGGFSFRF 43 RFamide neuropeptide QRF-amide NP05241 SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII 41 Sauvagine/corticotropin- Corticoliberin releasing factor/urotensin I NP05242 DNPSLSIDLTFHLLRTLLELARTQSQRERAEQNRIIFDSV 40 Sauvagine/corticotropin- Urocortin releasing factor/urotensin I NP05243 IVLSLDVPIGLLQILLEQARARAAREQATTNARILARVGHC 41 Sauvagine/corticotropin- Urocortin-2 releasing factor/urotensin I NP05244 FTLSLDVPTNIMNLLFNIAKAKNLRAQAAANAHLMAQI 38 Sauvagine/corticotropin- Urocortin-3 releasing factor/urotensin I NP05299 MDPNAAYVNMSNHHRGLASANVDFAFSLYKHLVALSPKKNIFISPVSISMALAMLS 383 Serpin Corticosteroid-binding LGTCGHTRAQLLQGLGFNLTERSETEIHQGFQHLHQLFAKSDTSLEMTMGNALFLD globulin GSLELLESFSADIKHYYESEVLAMNFQDWATASRQINSYVKNKTQGKIVDLFSGLD SPAILVLVNYIFFKGTWTQPFDLASTREENFYVDETTVVKVPMMLQSSTISYLHDSE LPCQLVQMNYVGNGTVFFILPDKGKMNTVIAALSRDTINRWSAGLTSSQVDLYIPK VTISGVYDLGDVLEEMGIADLFTNQANFSRITQDAQLKSSKVVHKAVLQLNEEGV DTAGSTGVTLNLTSKPIILRFNQPFIIMIFDHFTWSSLFLARVMNPV NP05300 QPLLAHGDKSLQGPQPPRHQLSEPAPAYHRITPTITNFALRLYKELAADAPGNIFFSP 403 Serpin Serpin A11 VSISTTLALLSLGAQANTSALILEGLGFNLTETPEADIHQGFRSLLHTLALPSPKLELK VGNSLFLDKRLKPRQHYLDSIKELYGAFAFSANFTDSVTTGRQINDYLRRQTYGQV VDCLPEFSQDTFMVLANYIFFKAKWKHPFSRYQTQKQESFFVDERTSLQVPMMHQ KEMHRFLYDQDLACTVLQIEYRGNALALLVLPDPGKMKQVEAALQPQTLRKWGQ LLLPSLLDLHLPRFSISGTYNLEDILPQIGLTNILNLEADFSGVTGQLNKTISKVSHKA MVDMSEKGTEAGAASGLLSQPPSLNTMSDPHAHFNRPFLLLLWEVTTQSLLFLGK VVNPVAG NP05301 LKPSFSPRNYKALSEVQGWKQRMAAKELARQNMDLGFKLLKKLAFYNPGRNIFLS 394 Serpin Serpin A12 PLSISTAFSMLCLGAQDSTLDEIKQGFNFRKMPEKDLHEGFHYIIHELTQKTQDLKL SIGNTLFIDQRLQPQRKFLEDAKNFYSAETILTNFQNLEMAQKQINDFISQKTHGKIN NLIENIDPGTVMLLANYIFFRARWKHEFDPNVTKEEDFFLEKNSSVKVPMMFRSGI YQVGYDDKLSCTILEIPYQKNITAIFILPDEGKLKHLEKGLQVDTFSRWKTLLSRRV VDVSVPRLHMTGTFDLKKTLSYIGVSKIFEEHGDLTKIAPHRSLKVGEAVHKAELK MDERGTEGAAGTGAQTLPMETPLVVKIDKPYLLLIYSEKIPSVLFLGKIVNPIGK NP05302 SRCSAQKNTEFAVDLYQEVSLSHKDNIIFSPLGITLVLEMVQLGAKGKAQQQIRQTL 387 Serpin Serpin I2 KQQETSAGEEFFVLKSFFSAISEKKQEFTFNLANALYLQEGFTVKEQYLHGNKEFFQ SAIKLVDFQDAKACAEMISTWVERKTDGKIKDMFSGEEFGPLTRLVLVNAIYFKGD WKQKFRKEDTQLINFTKKNGSTVKIPMMKALLRTKYGYFSESSLNYQVLELSYKG DEFSLIIILPAEGMDIEEVEKLITAQQILKWLSEMQEEEVEISLPRFKVEQKVDFKDV LYSLNITEIFSGGCDLSGITDSSEVYVSQVTQKVFFEINEDGSEAATSTGIHIPVIMSL AQSQFIANHPFLFIMKHNPTESILFMGRVTNPDTQEIKGRDLDSL NP05326 DRVY 4 Serpin Angiotensin 1-4 NP05327 DRVYI 5 Serpin Angiotensin 1-5 NP05328 DRVYIHP 7 Serpin Angiotensin 1-7 NP05329 DRVYIHPFH 9 Serpin Angiotensin 1-9 NP05330 DRVYIHPFHL 10 Serpin Angiotensin-1 NP05331 DRVYIHPF 8 Serpin Angiotensin-2 NP05332 RVYIHPF 7 Serpin Angiotensin-3 NP05333 VYIHPF 6 Serpin Angiotensin-4 NP05334 DRVYIHPFHLVIHNESTCEQLAKANAGKPKDPTFIPAPIQAKTSPVDEKALQDQLVL 452 Serpin Angiotensinogen VAAKLDTEDKLRAAMVGMLANFLGFRIYGMHSELWGVVHGATVLSPTAVFGTLA SLYLGALDHTADRLQAILGVPWKDKNCTSRLDAHKVLSALQAVQGLLVAQGRAD SQAQLLLSTVVGVFTAPGLHLKQPFVQGLALYTPVVLPRSLDFTELDVAAEKIDRF MQAVTGWKTGCSLMGASVDSTLAFNTYVHFQGKMKGFSLLAEPQEFWVDNSTSV SVPMLSGMGTFQHWSDIQDNFSVTQVPFTESACLLLIQPHYASDLDKVEGLTFQQN SLNWMKKLSPRTIHLTMPQLVLQGSYDLQDLLAQAELPAILHTELNLQKLSNDRIR VGEVLNSIFFELEADEREPTESTQQLNKPEVLEVTLNRPFLFAVYDQSATALHFLGR VANPLSTA NP05404 DRMPCRNFFWKTFSSCK 17 Somastostatin Cortistatin-17 NP05405 QEGAPPQQSARRDRMPCRNFFWKTFSSCK 29 Somastostatin Cortistatin-29 (Potential) NP05406 AGCKNFFWKTFTSC 14 Somastostatin Somatostatin-14 NP05407 SANSNPAMAPRERKAGCKNFFWKTFTSC 28 Somastostatin Somatostatin-28 NP05514 LPICPGGAARCQVTLRDLFDRAVVLSHYIHNLSSEMFSEFDKRYTHGRGFITKAINS 199 Somatotropin/prolactin Prolactin CHTSSLATPEDKEQAQQMNQKDFLSLIVSILRSWNEPLYHLVTEVRGMQEAPEAIL SKAVEIEEQTKRLLEGMELIVSQVHPETKENEIYPVWSGLPSLQMADEESRLSAYY NLLHCLRRDSHKIDNYLKLLKCRIIHNNNC NP05529 RPKPQFFGLM 10 Tachykinin Substance P NP05682 ALNSVAYERSAMQNYE 16 Tachykinin C-terminal-flanking peptide NP05683 HKTDSFVGLM 10 Tachykinin Neurokinin A NP05684 DADSSIEKQVALLKALYGHGQISHKRHKTDSFVGLM 36 Tachykinin Neuropeptide K NP05685 RPKPQQFFGLM 11 Tachykinin Substance P NP05686 DMHDFFVGLM 10 Tachykinin Neurokinin-B NP05799 QPEAAQQEAVTAAEHPGLDDFLRQVERLLFLRENIQRLQGDQGEHSASQIFQSDWL 218 TRH Pro-thyrotropin-releasing SKRQHPGKREEEEEEGVEEEEEEEGGAVGPHKRQHPGRREDEASWSVDVTQHKRQ hormone HPGRRSPWLAYAVPKRQHPGRRLADPKAQRSWEEEEEEEEREEDLMPEKRQHPGK RALGGPCGPQGAYGQAGLLLGLLDDLSRSQGAEEKRQHPGRRAAWVREPLEE NP05800 QHP 3 TRH Thyrotropin-releasing hormone NP05810 ETPDCFWKYCV 11 Urotensin-2 Urotensin-2 NP05811 ACFWKYCV 8 Urotensin-2 Urotensin-2B NP05890 AAPDLDVRKCLPCGPGGKGRCFGPNICCAEELGCFVGTAEALRCQEENYLPSPCQS 94 Vasopressin/oxytocin Neurophysin 1 GQKACGSGGRCAVLGLCCSPDGCHADPACDAEATFSQR NP05891 CYIQNCPLG 9 Vasopressin/oxytocin Oxytocin NP05892 CYFQNCPRG 9 Vasopressin/oxytocin Arg-vasopressin NP05893 ASDRSNATQLDGPAGALLLRLVQLAGAPEPFEPAQPDAY 39 Vasopressin/oxytocin Copeptin NP05894 AMSDLELRQCLPCGPGGKGRCFGPSICCADELGCFVGTAEALRCQEENYLPSPCQS 93 Vasopressin/oxytocin Neurophysin 2 GQKACGSGGRCAAFGVCCNDESCVTEPECREGFHRRA NP05912 TLQPPSALRRRHYHHALPPSRHYP 24 VGF Antimicrobial peptide VGF[554-577] NP05913 RPESALLGGSEAGERLLQQGLAQVEA 26 VGF Neuroendocrine regulatory peptide-1 NP05914 QAEATRQAAAQEERLADLASDLLLQYLLQGGARQRGLG 38 VGF Neuroendocrine regulatory peptide-2 NP05915 APPGRPEAQPPPLSSEHKEPVAGDAVPGPKDGSAPEVRGARNSEPQDEGELFQGVD 593 VGF Neurosecretory protein VGF PRALAAVLLQALDRPASPPAPSGSQQGPEEEAAEALLTETVRSQTHSLPAPESPEPA APPRPQTPENGPEASDPSEELEALASLLQELRDFSPSSAKRQQETAAAETETRTHTLT RVNLESPGPERVWRASWGEFQARVPERAPLPPPAPSQFQARMPDSGPLPETHKFGE GVSSPKTHLGEALAPLSKAYQGVAAPFPKARRPESALLGGSEAGERLLQQGLAQVE AGRRQAEATRQAAAQEERLADLASDLLLQYLLQGGARQRGLGGRGLQEAAEERE SAREEEEAEQERRGGEERVGEEDEEAAEAEAEAEEAERARQNALLFAEEEDGEAG AEDKRSQEETPGHRRKEAEGTEEGGEEEDDEEMDPQTIDSLIELSTKLHLPADDVVS IIEEVEEKRKRKKNAPPEPVPPPRAAPAPTHVRSPQPPPPAPAPARDELPDWNEVLPP WDREEDEVYPPGPYHPFPNYIRPRTLQPPSALRRRHYHHALPPSRHYPGREAQARR AQEEAEAEERRLQEQEELENYIEHVLLRRP
TABLE-US-00005 TABLE 5 Peptide Analogue suquence (potential glycan highlighted Peptide hormone name analogue name Trade name Producer in bold/underscore) (SEQ ID NO) Amylin NN9838/AM833 NA Novo Nordisk ? Amylin Pramlintide Symlin Bristol-Myers KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY- Squibb NH2 (SEQ ID NO: 280) ANP 32 Urodilatin Ularitide ? TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRY (SEQ ID NO: 281) ANP 28 Carperitide HANP Daiichi Sankyo SLRRSSCFGGRMDRIGAQSGLGCNSFRY (SEQ ID NO: 282) ANP 40 mutant ANP ? ? SLRRSSCFGGRMDRIGAQSGLGCNSFRYRITAREDKQ GWA (SEQ ID NO: 283) BNP 32 Nesiritide Natrecor Scios, Inc. SPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH (SEQ ID NO: 284) Bradykinin Icatibant Firazyr Shire RRPXGXSXXR (SEQ ID NO: 285) Calcitonin Salmon Calcitonin Fortical/ Several CSNLSTCVLGKLSQELHKLQTYPRTNTGSGTP (SEQ ID Miacalcin... NO: 286) DNP/CNP cdNP/cenderitide ? ? GLSKGCFGLKLDRIGSMSGLGCPSLRDPRPNAPSTSA (snake DNP CNP combination) (SEQ ID NO: 287) GIP/GLP-1 NN9277 GG dual NA Novo Nordisk ? agonist GIP/GLP-1/Glucagon NN9423 Triagonist NA Novo Nordisk ? 1706 GLP-1 Exenatide BYETTA/ Bristol-Myers HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPP BYDUREON Squibb S (SEQ ID NO: 288) GLP-1 Lixisenatide Lyxumia/Adlyxin Sanofi HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPS KKKKKK (SEQ ID NO: 289) GLP-1 GLP-1(7-37) linked to Albumin) Albiglutide Tanzeum GlaxoSmithKline ? GLP-1 GLP-1(7-37) linked to an Fc Dulaglutide Trulicity Eli Lilly ? fragment of human IgG4 GLP-1 LIRAGLUTIDE Victoza/Saxenda Novo Nordisk HAEGTFTSDVSSYLEGQAAK-E-EFIAWLVRGRG (SEQ ID NO: 290) GLP-1 (37 aa acylated peptide) Semaglutide NA Novo Nordisk HAEGTFTSDVSSYLEGQAAKEEFIIAWLVKGRG (SEQ ID NO: 291) GLP-2 Teduglutide GATTEX NPS HGDGSFSDEMNTILDNLAARDFINWLIQTKITD (SEQ Pharmaceuticals ID NO: 292) Glucagon NN9030/G530S NA Novo Nordisk ? Glucagon (produced in Saccharomyces) Glucagon GlucaGen Novo Nordisk HSQGTFTSDYSKYLDSRRAQDFVQWLMNT (SEQ ID NO: 293) Glucagon (produced in Escherichia coli) Glucagon Glucagon Eli Lilly HSQGTFTSDYSKYLDSRRAQDFVQWLMNT (SEQ ID NO: 293; same as previous line) PTHR Abaloparatide/ Tymlos Radiuspharm AVSEHQLLHDKGKSIQDLRRRELLEKLLXKLHTA BA058 (SEQ ID NO: 294) PYY NN9747/NN9748 NA Novo Nordisk Secretin Secretin Secremax/ Repligen Corp HSDGTFTSELSRLRDSARLQRLLQGLV (SEQ ID NO: Secreflow 295) Somatoliberin Sermorelin Sermorelin Emd serono inc YADAIFTNSYRKVLGQLSARKLLQDIMSRQ (SEQ ID acetate NO: 296) Somatoliberin Tesamorelin Egrifta Thera- YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQE technologies RGARARL (SEQ ID NO: 297)
TABLE-US-00006 TABLE 6 *(Bold; identified or conserved in human, Italic; identified in non-human and not conserved; the SEQ ID Nos. are found in the tables infra) *Site position (Bold; identified or conserved in human, Italic; Peptide identified in non- No. Entry_ Peptide Peptide Peptide Hormone human and not of Human Start Peptide_Sequence end Name Family conserved) sites P05408 200 SVPHFSDEDKDPE 212 C-terminal 7B2 S200 1 peptide (By similarity) P05408 27 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQA 212 Neuroendocrine 7B2 S28, T31, S36, 4 MNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAELTGDNIPK protein 7B2 T134 S200 DFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAEFSREFQ LHQHLFDPEHDYPGLGKWNKKLLYEKMKGGERRKRRSV NPYLQGQRLDNVVAKKSVPHFSDEDKDPE P05408 27 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQA 176 N-terminal 7B2 S28, T31, S36, 3 MNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAELTGDNIPK peptide (By T134 DFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAEFSREFQ similarity) LHQHLFDPEHDYPGLGKWNKKLLYEKMKGGE Q8N6N7 1 MALQADFDRAAEDVRKLKARPDDGELKELYGLYKQAIV 88 Acyl-CoA- ACBP GDINIACPGMLDLKGKAKWEAWNLKKGLSTEDATSAYIS binding domain- KAKELIEKYGI containing protein 7 P07108 2 SQAEFEKAAEEVRHLKTKPSDEEMLFIYGHYKQATVGDI 87 Acyl-CoA- ACBP T36 or T42 1 NTERPGMLDFTGKAKWDAWNELKGTSKEDAMKAYINK binding protein (ambiguous) VEELKKKYGI P35318 95 YRQSMNNFQGLRSEGCREGTCTVQKLAHQIYQFTDKDKD 146 Adrenomedullin Adrenomedullin NVAPRSKISPQGY Q7Z4H4 101 TQAQLLRVGCVLGTCQVQNLSHRLWQLMGPAGRQDSAP 147 Adrenomedullin- Adrenomedullin VDPSSPHSY 2 (By similarity) Q7Z4H4 108 VGCVLGTCQVQNLSHRLWQLMGPAGRQDSAPVDPSSPH 147 Intermedin-short Adrenomedullin SY (Potential) P35318 22 ARLDVASEFRKKWNKWALSR 41 Proadrenomedull Adrenomedullin in N-20 terminal peptide Q9ULZ1 65 QRPRLSHKGPMPF 77 Apelin-13 (By Apelin similarity) Q9ULZ1 50 NGPGPWQGGRRKFRRQRPRLSHKGPMPF 77 Apelin-28 (By Apelin similarity) Q9ULZ1 47 GSRNGPGPWQGGRRKFRRQRPRLSHKGPMPF 77 Apelin-31 (By Apelin similarity) Q9ULZ1 42 LVQPRGSRNGPGPWQGGRRKFRRQRPRLSHKGPMPF 77 Apelin-36 (By Apelin similarity) P07492 24 VPLPAGGGTVLTKMYPRGNHWAVGHLM 50 Gastrin-releasing Bombesin/neurom P27, T32 2 peptide edin-B/ranatensin P08949 47 GNLWATGHFM 56 Neuromedin-B Bombesin/neurom T52 0 edin-B/ranatensin P08949 25 APLSWDLPEPRSRASKIRVHSRGNLWATGHFM 56 Neuromedin-B- Bombesin/neurom T52 0 32 edin-B/ranatensin P07492 41 GNHWAVGHLM 50 Neuromedin-C Bombesin/neurom edin-B/ranatensin P01042 381 RPPGFSPFR 389 Bradykinin Bradykinin P01042 380 KRPPGFSPFR 389 Lysyl-bradykinin Bradykinin P01042 376 ISLMKRPPGFSPFR 389 T-kinin Bradykinin P01258 85 CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP 116 Calcitonin Calcitonin P06881 83 ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSKAF 119 Calcitonin gene- Calcitonin T91 1 related peptide 1 P10092 82 ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSKAF 118 Calcitonin gene- Calcitonin T90 1 related peptide 2 P10997 34 KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY 70 Islet amyloid Calcitonin T38, T42, T63 3 polypeptide P01258 121 DMSSDLERDHRPHVSMPQNAN 141 Katacalcin Calcitonin S135 0 Q16568 28 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKKLKS 66 CART(1-39) CART Q16568 69 VPIYEKKYGQVPMCDAGEQCAVRKGARIGKLCDCPRGTS 116 CART(42-89) CART CNSFLLKCL Q16568 28 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKKLKSK 116 Cocaine- and CART RVPIYEKKYGQVPMCDAGEQCAVRKGARIGKLCDCPRGT amphetamine- SCNSFLLKCL regulated P23435 58 GSAKVAFSAIRSTNH 72 [des-Ser1] Cerebellins cerebellin P23435 57 SGSAKVAFSAIRSTNH 72 Cerebellin Cerebellins P23435 22 QNEIEPIVLEGKCLVVCDSNPTSDPTGTALGISVRSGSAKV 193 Cerebellin-1 Cerebellins AFSAIRSTNHEPSEMSNRTMIIYEDQVLVNIGNNEDSERST FIAPRKGIYSENFHVVKVYNRQTIQVSLMLNGWPVISAFA GDQDVTREAASNGVLIQMEKGDRAYLKLERGNLMGGW KYSTFSGFLVFPL Q8IUK8 52 QNDIEPIVLEGKCLVVCDSSPSADGAVTSSLGISVRSGSAK 224 Cerebellin-2 Cerebellins VAFSATRSTNHEPSEMSNRTMTIYEDQVLVNIGNHFDLAS SIFVAPRKGIYSFSFHVVKVYNRQTIQVSLMQNGYPVISAF AGDQDVTREAASNGVLLLMEREDKVHLKLERGNLMGG WKYSTFSGFLVFPL Q6UW01 33 QEGSEPVLLEGECLVVCEPGRAAAGGPGGAALGEAPPGR 205 Cerebellin-3 Cerebellins VAFAAVRSHHHEPAGETGNGTSGAIYEDQVLVNEGGGED RASGSFVAPVRGVYSFRFHVVKVYNRQTVQVSLMLNTW PVISAFANDPDVTREAATSSVLLPLDPGDRVSLRLRRGNL LGGWKYSSFSGFLIFPL Q9NTU7 28 QNDIEPIVLEGKCLVVCDSNPATDSKGSSSSPLGISVRAAN 201 Cerebellin-4 Cerebellins SKVAFSAVRSTNHEPSEMSNKTRIIYEDQILVNVGNEFTLE SVEVAPRKGIYSFSFHVIKVYQSQTIQVNLMLNGKPVISAF AGDKDVTREAATNGVLLYLDKEDKVYLKLEKGNLVGG WQYSTFSGFLVFPL P10645 380 AYGFRGPGPQL 390 AL-11 Chromogranin/ secretogranin P05060 617 SAEFPDFYDSEEPSVTHQEAENEKDRADQTVLTEDEKKEL 673 CCB peptide Chromogranin/ S631 1 ENLAAMDLELQKIAEKF secretogranin P10645 19 LPVNSPMNKGDIEVMKCIVEVISDTLSKPSPMPVSQECFE 457 Chromogranin-A Chromogranin/ S45, S48, S54, 39 TLRGDERILSILRHQNLLKELQDLALQGAKERAHQQKKHS secretogranin S98, E122, S126, GFEDELSEVLENQSSQAELKEAVEEPSSKDVMEKREDSKE A145, T146, AEKSGEATDGARPQALPEPMQESKAEGNNQAPGEEEEEEE A153, L154, EATNTHPPASLPSQKYPGPQAEGDSEGLSQGLVDREKGLS S161, A169, AEPGWQAKREEEEEEEEEAEAGEEAVPEEEGPTVVLNPHP N182, T183, SLGYKEIRKGESRSEALAVDGAGKPGAEEAQDPEGKGEQ P185, A187, EHSQQKEEEEEMAVVPQGLFRGGKSGELEQEEERLSKEW S188, S191, EDSKRWSKMDQLAKELTAEKRLEGQEEEEDNRDSSMKL E200, L206, SFRARAYGFRGPGPQLRRGWRPSSREDSLEAGLPLQVRGY S207, V211, PEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQLQAL S218, T251 RRG L254, N255, P256, S259, Y262, P283, E286, P291, S300, M309, V312, T353, A378, S398, G413 P10645 134 EDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQAPGE 225 EA-92 Chromogranin/ A145, T146, 17 EEEEEEEATNTHPPASLPSQKYPGPQAEGDSEGLSQGLVD secretogranin A153, L154, REKGLSAEPGWQA S161, A169, N182, T183, P185, A187, S188, S191, E200, L206, S207, V211, S218 P10645 420 EEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRR 456 ER-37 Chromogranin/ S436 or 1 secretogranin S438(Ambiguous) P10645 228 EEEEEEEEEAEAGEEAVPEEEGPTVVLNPHPSL 260 ES-43 Chromogranin/ T251, L254, 5 secretogranin N255, P256, S259 P05060 440 FLGEGHHRVQENQMDKARRHPQGAWKELDRNYLNYGE 513 GAWK peptide Chromogranin/ H446, N451, 5 EGAPGKWQQQGDLQDTKENREEARFQDKQYSSHHTAE secretogranin Y474, T511, A512 P10645 413 GYPEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQLQ 456 GR-44 Chromogranin/ G413 1 ALRR secretogranin P10645 393 GWRPSSREDSLEAGLPLQV 411 GV-19 Chromogranin/ 5398 1 secretogranin P10645 358 LEGQEEEEDNRDSSMKLSF 376 LF-19 Chromogranin/ secretogranin P10645 272 SEALAVDGAGKPGALEAQDPEGKGEQEHSQQKEEEEEM 319 Pancreastatin Chromogranin/ P283, E286, 6 AVVPQGLFRG secretogranin P291, S300, M309, V312 P05060 21 MPVDNRNHNEGMVTRCIIEVLSNALSKSSAPPITPECRQV 677 Secretogranin-1 Chromogranin/ T34, S46, S48, 40 LKTSRKDVKDKETFENENTKFEVRLLRDPADASEAHESSS secretogranin S49, P52, T79, RGEAGAPGEEDIQGPTKADILKWAEGGGHSRERADEPQ S93, A95, S100, WSLYPSDSQVSEEVKTRHSEKSQREDEEEEEGENYQKGE A104, E109, RGEDSSEEKHLEEPGETQNAFLNERKQASAIKKEELVARS T116, S140, ETHAAGHSQEKTHSREKSSQESGEETGSQENHPQESKGQP S144, S149, RSQEESEEGEEDATSEVDKRRTRPRHHHGRSRPDRSSQGG T194, N196 , SLPSEEKGHPQEESEESNVSMASLGEKRDHHSTHYRASEE S206, S293, EPEYGEEIKGYPGVQAPEDLEWERYRGRGSEEYRAPRPQS S294, S298, EESWDEEDKRNYPSLELDKMAHGYGEESEEERGLEPGKG P307, S317, RHHRGRGGEPRAYFMSDTREEKRFLGEGHHRVQENQMD M318, S320, KARRHPQGAWKELDRNYLNYGEEGAPGKWQQQGDLQD D326, T330, TKENREEARFQDKQYSSHHTAEKRKRLGELFNPYYDPLQ Y348, P349, WKSSHFERRDNMNDNFLEGEEENELTLNEKNFFPEYNYD G350, K396, WVVEKKPFSEDVNWGYEKRNLARVPKLDLKRQYDRVAQ M397, H446, LDQLLHYRKKSAEFPDFYDSEEPVSTHQEAENEKDRADQ Y474, T511, TVLTEDEKKELENLAAMDLELQKIAEKFSQRG A512, T556 S577, N588, S631 P13521 31 QRNQLLQKEPDLRLENVQKFPSPEMIRALEYIENLRQQAH 617 Secretogranin-2 Chromogranin/ S52, S75, A136, 16 KEESSPDYNPYQGVSVPLQQKENGDESHLPERDSLSEED secretogranin Y190, T191, WMRIILEALRQAENEPQSAPKENKPYALNSEKNFPMDMS S194, T197, DDYETQQWPERKLKHMQFPPMYEENSRDNPFKRTNEIVE S259, T261, EQYTPQSLATLESVFQELGKLTGPNNQKRERMDEEQKLY S378, T450, TDDEDDIYKANNIAYEDVVGGEDWNPVEEKIESQTQEEV N489, G529, RDSKENIEKNEQINDEMKRSGQLGIQEEDLRKESKDQLSD S556, S566, DVSKVIAYLKRLVNAAGSGRLQNGQNGERATRLFEKPLD P573 SQSIYQLIEISRNLQIPPEDLIEMLKTGEKPNGSVEPERELD LPVDLDDISEADLDHPDLFQNRMLSKSGYPKTPGRAGILA LPDGLSVEDILNLLGMESAANQKTSYFPNPYNQEKVLPRL PYGAGRSRSNQLPKAAWIPHVENRQMAYENLNDKDQEL GEYLARMLVKYPEIINSNQVKRVPGQGSSEDDLQEEEQIE QAIKEHLNQGSSQETDKLAPVSKRFPVGPPKNDDTPNRQ YVVDEDLLMKVLEYLNQEKAEKGREHIAKRAMENM Q8WXD2 20 FPKPGGSQDKSLHNRELSAERPLNEQIAEAEEDKIKKTYPP 468 Secretogranin-3 Chromogranin/ P60, T82, S122, 15 ENKPGQSNYSFVDNLNLLKAITEKEKIEKERQSIRSSPLDN secretogranin T123, T144, KLNVEDVDSTKNRKLIDDYDSTKSGLDHKFQDDPDGLHQ A157, D211, LDGTPLTAEDIVHKIAARIYEENDRAVFDKIVSKLLNLGLI P212, T216, FESQAHTLEDEVAEVLQKLISKEANNYEEDPNKPTSWTE S217, T219, NQAGKIPEKVTPMAAIQDGLAKGENDETVSNTLTLTNGL T231, I236, ERRTKTYSEDNFLELQYFPNFYALLKSIDSEKEAKEKETLI A241, S359 TIMKTLIDFVKMMVKYGTISPEEGVSYLENLDEMIALQTK NKLEKNATDNISKLFPAPSEKSHEETDSTKEEAAKMEKEY GSLKDSTKDDNSNPGGKTDEPKGKTEAYLEAIRKNIEWL KKHDKKGNKEDYDLSKMRDFINKQADAYVEKGILDKEE AEAIKRIYSSL P13521 182 TNEIVEEQYTPQSLATLESVFQELGKLTGPNNQ 214 Secretoneurin Chromogranin/ Y190, T191, 4 secretogranin S194, T197 P10645 322 SGELEQEEERLSKEWEDS 339 SS-18 Chromogranin/ secretogranin P10645 19 LPVNSPMNKGDIEVMKCIVEVISDTLSKPSPMPVSQECFE 94 Vasostatin-1 Chromogranin/ S45, S48, S54 3 TLRGDERILSILRHQNLLKELQDLALQGAKERAHQQ secretogranin P10645 19 LPVNSPMNKGDIEVMKCIVEVISDTLSKPSPMPVSQECFE 131 Vasostatin-2 Chromogranin/ S45, S48, S54, 6 TLRGDERILSILRHQNLLKELQDLALQGAKERAHQQKKHS secretogranin S98, E122, S126 GFEDELSEVLENQSSQAELKEAVEEPSSKDVME P10645 342 WSKMDQLA 349 WA-8 Chromogranin/ secretogranin P10645 342 WSKMDQLAKELTAE 355 WE-14 Chromogranin/ T353 1 secretogranin P01042 19 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFVLY 644 Kininogen-1 Cystatin T137, T151, 15 RITEATKTVGSDTFYSFKYEIKEGDCPVQSGKTWQDCEYK T261, T273, DAAKAATGECTATVGKRSSTKFSVATQTCQITPAEGPVV E399, T400, TAQYDCLGCVHPISTQSPDLEPILRHGIQYFNNNTQHSSLF T401, S403, MLNEVKRAQRQVVAGLNFRITYSIVQTNCSKENFLFLTPD T407, S408, CKSLWNGDTGECTDNAYIDIQLRIASFSQNCDIYPGKDEV T546, S604, QPPTKICVGCPRDIPTNSPELEETLTHTITKLNAENNATFYF T610, S611, KIDNVKKARVQVVAGKKYFIDFVARETTCSKESNEELTES T628 CETKKLGQSLDCNAEVYVVPWEKKIYPTVNCQPLGMISL MKRPPGESPERSSRIGEIKEETTVSPPHTSMAPAQDEERDS GKEQGHTRRHDWGHEKQRKHNLGHGHKHERDQGHGHQ RGHGLGHGHEQQHGLGHGHKFKLDDDLEHQGGHVLDH GHKHKHGHGHGKHKNKGKKNGKHNGWKTEHLASSSED STTPSAQTQEKTEGPTPIPSLAKPGVTVTFSDFQDSDLIAT MMPPISPAPIQSDDDWIPDIQIDPNGLSFNPISDFPDTTSPK CPGRPWKSVSEINPTTQMKESYYFDLTDGLS P01042 19 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFVLY 380 Kininogen-1 Cystatin T137, 4 RITEATKTVGSDTFYSFKYEIKEGDCPVQSGKTWQDCEYK heavy chain T151, T261, DAAKAATGECTATVGKRSSTKFSVATQTCQITPAEGPVV T273 TAQYDCLGCVHPISTQSPDLEPILRHGIQYFNNNTQHSSLF MLNEVKRAQRQVVAGLNFRITYSIVQTNCSKENFLFLTPD CKSLWNGDTGECTDNAYIDIQLRIASFSQNCDIYPGKDEV QPPTKICVGCPRDIPTNSPELEETLTHTITKLNAENNATFYF KIDNVKKARVQVVAGKKYFIDFVARETTCSKESNEELTES CETKKLGQSLDCNAEVYVVPWEKKIYPTVNCQPLGMISL MK P01042 390 SSRIGEIKEETTVSPPHTSMAPAQDEERDSGKEQGHTRRH 644 Kininogen-1 Cystatin E399, T400, 11 DWGHEKQRKHNLGHGHKHERDQGHGHQRGHGLGHGHE light chain T401, S403, QQHGLGHGHKFKLDDDLEHQGGHVLDHGHKHKHGHGH T407, S408, GKHKNKGKKNGKHNGWKTEHLASSSEDSTTPSAQTQEK T546, S604, FEGPTPIPSLAKPGVTVTFSDFQDSDLIATMMPPISPAPIQS T610, S611, DDDWIPDIQIDPNGLSFNPISDFPDTTSPKCPGRPWKSVSEI T628 NPTTQMKESYYFDLTDGLS P01042 431 WGHE 434 Low molecular Cystatin weight growth- promoting factor P05305 53 CSCSSLMDKECVYFCHLDIIWVNTPEHVVPYGLGSPRS 90 Big endothelin-1 Endothelin/ sarafotoxin P05305 53 CSCSSLMDKECVYFCHLDIIW 73 Endothelin-1 Endothelin/ sarafotoxin P20800 49 CSCSSWLDKECVYFCHLDIIW 69 Endothelin-2 Endothelin/ sarafotoxin P14138 97 CTCFTYKDKECVYYCHLDIIW 117 Endothelin-3 Endothelin/ sarafotoxin O15130 93 AGEGLNSQFWSLAAPQRF 110 Neuropeptide AF FMRFamide related peptide O15130 69 FLFQPQRF 76 Neuropeptide FF FMRFamide related peptide Q9HCQ7 56 SLNFEELKDWGPKNVIKMSTPAVNKMPHSFANLPLRF 92 Neuropeptide FMRFamide NPSF (Potential) related peptide Q9HCQ7 124 VPNLPQRF 131 Neuropeptide FMRFamide NPVF related peptide Q9HCQ7 81 MPHSFANLPLRF 92 Neuropeptide FMRFamide RFRP-1 related peptide Q9HCQ7 101 SAGATANLPLRS 112 Neuropeptide FMRFamide RFRP-2 related peptide (Potential) O15130 66 SQAFLFQPQRF 76 Neuropeptide SF FMRFamide related peptide P81277 34 TPDINPAWYASRGIRPVGRF 53 Prolactin- FMRFamide releasing peptide related peptide PrRP20 P81277 23 SRTHRHSMEIRTPDINPAWYASRGIRPVGRF 53 Prolactin- FMRFamide releasing peptide related peptide PrRP31 P22466 33 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 62 Galanin Galanin S55 0 P22466 65 ELRPEDDMKPGSFDRSIPENNIMRTHEFLSELHLKEAGALD 123 Galanin Galanin A113 1 RLLDLPAAASSEDIERS message- associated peptide Q9UBC7 25 APAHRGRGGWTLNSAGYLLGPVLHLPQMGDQDGKRETA 84 Galanin-like Galanin LEILDLWKAIDGLPYSHPPQPS peptide P01350 59 QLGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF 92 Big gastrin Gastrin/ cholecystokinin P06307 21 QPVPPADPAGSGLQRAEEAPRRQLRVSQRTDGESRAHLG 115 Cholecystokinin Gastrin/ A28, T50, N85 3 ALLARYIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYM cholecystokinin GWMDFGRRSAEEYEYPS P06307 92 ISDRDYMGWMDF 103 Cholecystokinin- Gastrin/ 12 cholecystokinin P06307 86 LDPSHRISDRDYMGWMDF 103 Cholecystokinin- Gastrin/ 18 (By cholecystokinin similarity) P06307 79 IVKNLQNLDPSHRISDRDYMGWMDF 103 Cholecystokinin- Gastrin/ N85 1 25 (By cholecystokinin similarity) P06307 71 KAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 103 Cholecystokinin- Gastrin/ N85 1 33 cholecystokinin P06307 65 YIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 103 Cholecystokinin- Gastrin/ N85 1 39 cholecystokinin P06307 99 GWMDF 103 Cholecystokinin- Gastrin/ 5 (By similarity) cholecystokinin P06307 46 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKNLQ 103 Cholecystokinin- Gastrin/ T50, N85 2 NLDPSHRISDRDYMGWMDF 58 cholecystokinin P06307 46 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKNLQ 94 Cholecystokinin- Gastrin/ T50, N85 2 NLDPSHRISD 58 cholecystokinin desnonopeptide (By similarity) P06307 97 YMGWMDF 103 Cholecystokinin- Gastrin/ 7 (By similarity) cholecystokinin P06307 96 DYMGWMDF 103 Cholecystokinin- Gastrin/ 8 cholecystokinin P01350 76 QGPWLEEEEEAYGWMDF 92 Gastrin Gastrin/ cholecystokinin P01350 79 WLEEEEEAYGWMDF 92 Gastrin-14 Gastrin/ cholecystokinin P01350 41 DLELPWLEQQGPASHHRRQLGPQGPPHLVADPSKKQGPW 92 Gastrin-52 Gastrin/ LEEEEEAYGWMDF cholecystokinin P01350 87 YGWMDF 92 Gastrin-6 Gastrin/ cholecystokinin P01350 22 SWKPRSQQPDAPLGTGANRDLELPWLEQQGPASHHRRQL 92 Gastrin-71 Gastrin/ L33 or G34 1 GPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF cholecystokinin (Ambiguous) P09681 52 YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKH 93 Gastric inhibitory Glucagon NITQ polypeptide P01275 21 RSLQDTEEKSRSFSASQADPLSDPDQMNEDKRHSQGTFTS 89 Glicentin (By Glucagon S32, T59 2 DYSKYLDSRRAQDFVQWLMNTKRNRNNIA similarity) P01275 21 RSLQDTEEKSRSFSASQADPLSDPDQMNED 50 Glicentin-related Glucagon S32 1 polypeptide (By similarity) P01275 53 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 81 Glucagon Glucagon T59 1 P01275 92 HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 128 Glucagon-like Glucagon T102 or T104 or 1 peptide 1 S105 (Ambiguous) P01275 98 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 128 Glucagon-like Glucagon T102 or T104 or 1 peptide 1(7-36) S105 (Ambiguous) P01275 98 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 127 Glucagon-like Glucagon T102 or T104 or 1 peptide 1(7-37) S105 (Ambiguous) P01275 146 HADGSFSDEMNTILDNLAARDFINVVLIQTKITD 178 Glucagon-like Glucagon peptide 2 (By similarity) P01282 81 HADGVFTSDFSKLLGQLSAKKYLESLM 107 Intestinal peptide Glucagon T87, S88 2 PHM-27 P01282 81 HADGVFTSDFSKLLGQLSAKKYLESLMGKRVSSNISEDPV 122 Intestinal peptide Glucagon T87, S88 2 PV PHV-42 P01275 53 HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA 89 Oxyntomodulin Glucagon T59 1 (By similarity) P18509 82 DVAHGILNEAYRKVLDQLSAGKHLQSLVARGVGGSLGG 129 PACAP-related Glucagon GAGDDAEPLS peptide P18509 132 HSDGIFTDSYSRYRKQMAVKKYLAAVL 158 Pituitary Glucagon adenylate cyclase- activating polypeptide 27 P18509 132 HSDGIFTDSYSRYRKQMAVKKYLAAVLGKRYKQRVKNK 169 Pituitary Glucagon adenylate cyclase- activating polypeptide 38 P09683 28 HSDGTFTSELSRLREGARLQRLLQGLV 54 Secretin Glucagon T34, S35, S38 3 P01286 32 YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERG 75 Somatoliberin Glucagon ARARL P01282 125 HSDAVFTDNYTRLRKQMAVKKYLNSILN 152 Vasoactive Glucagon T131 1 intestinal peptide P01148 37 DAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLRDLK 92 GnRH-associated GnRH T64 1 GALESLIEEETGQKKI peptide 1 O43555 37 ALS SAQDPQNALRPPGRALDTAAGSPVQTAHGLPSDALA 120 GnRH-associated GnRH PLDDSMPWEGRTTAQWSLHRKRHLARTLLTAAREPRPAP peptide 2 PSSNKV P01148 24 QHWSYGLRPG 33 Gonadoliberin-1 GnRH O43555 24 QHWSHGWYPG 33 Gonadoliberin-2 GnRH P01148 24 QHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFE 92 Progonadoliberin- GnRH T64 1 CTTHQPRSPLRDLKGALESLIEEETGQKKI 1 O43555 24 QHWSHGWYPGGKRALSSAQDPQNALRPPGRALDTAAGS 120 Progonadoliberin- GnRH PVQTAHGLPSDALAPLDDSMPWEGRTTAQWSLHRKRHL 2 ARTLLTAAREPRPAPPSSNKV P01308 90 GIVEQCCTSICSLYQLENYCN 110 Insulin A chain Insulin P01308 25 FVNQHLCGSHLVEALYLVCGERGFFYTPKT 54 Insulin B chain Insulin T51 1 (Ambiguous) P05019 49 GPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSRRAP 118 Insulin-like Insulin QTGIVDECCFRSCDLRRLEMYCAPLKPAKSA growth factor I P01344 25 AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSR 91 Insulin-like Insulin S74, T86 0 GIVEECCFRSCDLALLETYCATPAKSE growth factor II P01344 26 YRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRG 91 Insulin-like Insulin S74, T86 0 IVEECCFRSCDLALLETYCATPAKSE growth factor II Ala-25 Del P01344 93 DVSTPPTVLPDNFPRYPVGKFFQYDTVVKQSTQRL 126 Preptin Insulin S95, T96, T99 3 P04090 162 QLYSALANKCCHVGCTKRSLARFC 185 Relaxin A chain Insulin P04808 163 PYVALFEKCCLIGCTKRSLAKYC 185 Relaxin A chain Insulin (By similarity) P04090 25 DSWMEEVIKLCGRELVRAQIAICGMSTWS 53 Relaxin B chain Insulin P04808 23 VAAKWKDDVIKLCGRELVRAQIAICGMSTWS 53 Relaxin B chain Insulin (By similarity) Q8WXF3 119 DVLAGLSSSCCKWGCSKSEISSLC 142 Relaxin-3 A Insulin chain (By similarity) Q8WXF3 26 RAAPYGVRLCGREFIRAVIFTCGGSRW 52 Relaxin-3 B Insulin chain (By similarity) Q15726 112 YNWNSFGLRF 121 Kisspeptin-10 KISS1 Q15726 109 LPNYNVVNSFGLRF 121 Kisspeptin-13 KISS1 Q15726 108 DLPNYNWNSFGLRF 121 Kisspeptin-14 KISS1 Q15726 20 EPLEKVASVGNSRPTGQQLESLGLLAPGEQSLPCTERKPA 138 Metastasis- KISS1 A88 1 ATARLSRRGTSLSPPPESSGSPQQPGLSAPHSRQIPAPQGA suppressor KiSS- VLVQREKDLPNYNWNSFGLRFGKREAAPGNHGRSAGRG 1 Q15726 68 GTSLSPPPESSGSPQQPGLSAPHSRQIPAPQGAVLVQREKD 121 Metastin KISS1 A88 1 LPNYNVVNSFGLRF P41159 22 VPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVTGLDFI 167 Leptin Leptin PGLHPILTLSKMDQTLAVYQQILTSMPSRNVIQISNDLENL RDLLHVLAFSKSCHLPWASGLETLDSLGGVLEASGYSFEV VALSRLQGSLQDMLWQLDLSPGC P20382 147 DFDMLRCMLGRVYRPCWQV 165 Melanin- Melanin- concentrating concentrating hormone hormone P20382 131 EIGDEENSAKFPI 143 Neuropeptide- Melanin- glutamic acid- concentrating isoleucine hormone P20382 110 GSVAFPAENGVQNTESTQE 128 Neuropeptide- Melanin- T123, T125, 3 glycine-glutamic concentrating T126 acid(Potential) hormone P20382 22 ILLSASKSIRNLDDDMVFNTFRLGKGFQKEDTAEKSVIAPS 165 Pro-MCH Melanin- S57, S62, N93, 6 LEQYKNDESSFMNEEENKVSKNTGSKHNFLNHGLPLNLAI concentrating T123, T125 KPYLALKGSVAFPAENGVQNTESTQEKREIGDEENSAKFP hormone T126 IGRRDFDMLRCMLGRVYRPCWQV Q9UBU3 24 GSSFLSPEHQRVQQRKESKKPPAKLQP 50 Ghrelins-27 Motilin Q9UBU3 24 GSSFLSPEHQRVQQRKESKKPPAKLQPR 51 Ghrelins-28 Motilin P12872 26 FVPIFTYGELQRMQEKERNKGQ 47 Motilin Motilin P12872 50 SLSVWQRSGEEGPVDPAEPIREEENEMIKLTAPLEIGMRM 115 Motilin- Motilin NSRQLEKYPATLEGLLSEMLPQHAAK associated peptide Q9UBU3 76 FNAPFDVGIKLSGVQYQQHSQAL 98 Obestatin Motilin P12872 26 FVPIFTYGELQRMQEKERNKGQKKSLSVWQRSGEEGPVD 115 Promotilin Motilin PAEPIREEENEMIKLTAPLEIGMRMNSRQLEKYPATLEGLL SEMLPQHAAK Q15848 19 ETTTQGPGVLLPLPKGACTGWMAGIPGHPGHNGAPGRDG 244 Adiponectin NA T22 1 RDGTPGEKGEKGDPGLIGPKGDIGETGVPGAEGPRGFPGI QGRKGEPGEGAYVYRSAFSVGLETYVTIPNMPIRFTKIFY NQQNHYDGSTGKFHCNIPGLYYFAYHITVYMKDVKVSLF KKDKAMLFTYDQYQENNVDQASGSVLLHLEVGDQVWL QVYGEGERNGLYADNDNDSTFTGFLLYHDTN O00253 21 AQMGLAPMEGIRRPDQALLPELPGLGLRAPLKKTTAEQA 131 Agouti-related NA T55 1 EEDLLQEAQALAEVLDLQDREPRSSRRCVRLHESCLGQQ protein VPCCDPCATCYCRFFNAFCYCRKLGTAMNPCSRT Q9BZL1 1 MIEVVCNDRLGKKVRVKCNTDDTIGDLKKLIAAQTGTRW 73 Ubiquitin-like NA NKIVLKKWYTIFKDHVSLGDYEIHDGMNLELYYQ protein 5 P43490 1 MNPAAEAEFNILLATDSYKVTHYKQYPPNTSKVYSYFEC 491 Nicotinamide NAPRTase REKKFENSKLRKVKYEETVEYGLQVILNKYLKGKVVTKE phosphoribosyltr KIQEAKDVYKEHFQDDVFNEKGWNYILEKYDGHLPIEIK ansferase AVPEGFVIPRGNVLFTVENTDPECYWLTNVVIETILVQSWY PITVATNSREQKKILAKYLLETSGNLDGLEYKLHDFGYRG VSSQETAGIGASAHLVNFKGTDTVAGLALIKKYYGTKDP VPGYSVPAAEHSTITAWGKDHEKDAFEHIVTQFSSVPVSV VSDSYDIYNACEKIWGEDLRHLIVSRSTQAPLIIRPDSGNP LDTVLKVLEILGKKFPVTENSKGYKLLPPYLRVIQGDGVD INTLQEIVEGMKQKMVVSIENIAFGSGGGLLQKLTRDLLNC SFKCSYVVTNGLGINVFKDPVADPNKRSKKGRLSLHRTPA GNFVTLEEGKGDLEEYGQDLLHTVFKNGKVTKSYSFDEI RKNAQLNIELEAAHH P01160 124 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 151 Atrial natriuretic Natriuretic peptide S142, S148 2 factor P16860 103 SPKMVQGSGCFGRKMDRISSSSGLGCKV 130 BNP(1-28) Natriuretic peptide P16860 103 SPKMVQGSGCFGRKMDRISSSSGLGCKVL 131 BNP(1-29) Natriuretic peptide P16860 103 SPKMVQGSGCFGRKMDRISSSSGLGCKVLR 132 BNP(1-30) Natriuretic peptide P16860 104 PKMVQGSGCFGRKMDRISSSSGLGCKVLRR 133 BNP(2-31) Natriuretic peptide P16860 105 KMVQGSGCFGRKMDRISSSSGLGCKVL 131 BNP(3-29) Natriuretic peptide P16860 105 KMVQGSGCFGRKMDRISSSSGLGCKVLR 132 BNP(3-30) Natriuretic peptide P16860 105 KMVQGSGCFGRKMDRISSSSGLGCKVLRRH 134 BNP(3-32) Natriuretic peptide P16860 106 MVQGSGCFGRKMDRISSSSGLGCK 129 BNP(4-27) Natriuretic peptide P16860 106 MVQGSGCFGRKMDRISSSSGLGCKVL 131 BNP(4-29) Natriuretic peptide P16860 106 MVQGSGCFGRKMDRISSSSGLGCKVLR 132 BNP(4-30) Natriuretic peptide P16860 106 MVQGSGCFGRKMDRISSSSGLGCKVLRR 133 BNP(4-31) Natriuretic peptide P16860 106 MVQGSGCFGRKMDRISSSSGLGCKVLRRH 134 BNP(4-32) Natriuretic peptide P16860 107 VQGSGCFGRKMDRISSSSGLGCKVL 131 BNP(5-29) Natriuretic peptide P16860 107 VQGSGCFGRKMDRISSSSGLGCKVLRR 133 BNP(5-31) Natriuretic peptide P16860 107 VQGSGCFGRKMDRISSSSGLGCKVLRRH 134 BNP(5-32) Natriuretic peptide P16860 103 SPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH 134 Brain natriuretic Natriuretic peptide peptide 32 P01160 26 NPMYNAVSNADLMDFKNLLDHLEEKMPLED 55 Cardiodilatin- Natriuretic peptide Y29, A31, S33 1 related peptide (Ambiguous) P23582 105 GLSKGCFGLKLDRIGSMSGLGC 126 CNP-22 Natriuretic peptide P23582 98 YKGANKKGLSKGCFGLKLDRIGSMSGLGC 126 CNP-29 Natriuretic peptide P23582 74 DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCF 126 CNP-53 Natriuretic peptide GLKLDRIGSMSGLGC P16860 27 HPLGSPGSASDLETSGLQEQRNHLQGKLSELQVEQTSLEP 134 Natriuretic Natriuretic peptide T74, E85 2 LQESPRPTGVWKSREVATEGIRGHRKMVLYTLRAPRSPK peptides B MVQGSGCFGRKMDRISSSSGLGCKVLRRH P58417 22 ANLTNGGKSELLKSGSSKSTLKHIW1ESSKDLSISRLLSQT 271 Neurexophilin-1 Neurexophilin FRGKENDTDLDLRYDTPEPYSEQDLWDWLRNSTDLQEPR PRAKRRPIVKTGKFKKMFGWGDFHSNIKTVKLNLLITGKI VDHGNGTFSVYFRHNSTGQGNVSVSLVPPTKIVEFDLAQ QTVIDAKDSKSFNCRIEYEKVDKATKNTLCNYDPSKTCY QEQTQSHVSWLCSKPFKVICIYISFYSTDYKLVQKVCPDY NYHSDTPYFPSG O95156 23 KEVVHATEGLDWEDKDAPGTLVGNVVHSRIISPLRLFVK 264 Neurexophilin-2 Neurexophilin T29 1 QSPVPKPGPMAYADSMENFWDWLANIFEIQEPLARTKRR PIVKTGKFKKMFGWGDFHSNIKTVKLNLLITGKIVDHGNG TFSVYFRHNSTGLGNVSVSLVPPSKVVEFEVSPQSTLETKE SKSFNCRIEYEKTDRAKKTALCNFDPSKICYQEQTQSHVS WLCSKPFKVICIYIAFYSVDYKLVQKVCPDYNYHSETPYL SSG O95157 23 QDDGPPGSEDPERDDHEGQPRPRVPRKRGHISPKSRPMAN 252 Neurexophilin-3 Neurexophilin STLLGLLAPPGEAWGILGQPPNRPNHSPPPSAKVKKIFGW GDFYSNIKTVALNLLVTGKIVDHGNGTFSVHFQHNATGQ GNISISLVPPSKAVEFHQEQQIFIEAKASKIFNCRMEWEKV ERGRRTSLCTHDPAKICSRDHAQSSATVVSCSQPFKVVCVY IAFYSTDYRLVQKVCPDYNYHSDTPYYPSG O95158 24 QIPESGRPQYLGLRPAAAGAGAPGQQLPEPRSSDGLGVGR 308 Neurexophilin-4 Neurexophilin AWSWAWPTNHTGALARAGAAGALPAQRTKRKPSIKAAR AKKIFGWGDFYFRVHTLKFSLLVTGKIVDHVNGTFSVYFR HNSSSLGNLSVSIVPPSKRVEFGGVWLPGPVPHPLQSTLAL EGVLPGLGPPLGMAAAAAGPGLGGSLGGALAGPLGGAL GVPGAKESRAFNCHVEYEKTNRARKHRPCLYDPSQVCFT EHTQSQAAWLCAKPFKVICIFVSFLSFDYKLVQKVCPDYN FQSEHPYFG Q5H8A3 109 ILQRGSGTAAVDFTKKDHTATWGRPFFLFRPRN 141 Neuromedin-S Neuromedins P48645 142 FRVDEEFQSPFASQSRGYFLFRPRN 166 Neuromedin-U- Neuromedins 25 Q8NG41 25 WYKPAAGHSSYSVGRAAGLLSGLR 48 Neuropeptide B- Neuromedins 23 Q8NG41 25 WYKPAAGHSSYSVGRAAGLLSGLRRSPYA 53 Neuropeptide B- Neuromedins 29 P0C0P6 70 SFRNGVGTGMKKTSFQRAKS 89 Neuropeptide S Neuromedins Q8N729 33 WYKHVASPRYHTVGRAAGLLMGL 55 Neuropeptide W- Neuromedins 23 Q8N729 33 WYKHVASPRYHTVGRAAGLLMGLRRSPYLW 62 Neuropeptide W- Neuromedins 30 P30990 24 SDSEEEMKALEADFLTNMHTSKISKAHVPSWKMTLLNVC 148 Large Neurotensin SLVNNLNSPAEETGEVHEEELVARRKLPTALDGFSLEAML neuromedin N TIYQLHKICHSRAFQHWELIQEDILDTGNDKNGKEEVIKR KIPYIL P30990 144 IPYIL 148 Neuromedin N Neurotensin P30990 151 QLYENKPRRPYIL 163 Neurotensin Neurotensin P30990 166 DSYYY 170 Tail peptide Neurotensin (Potential) P01303 68 SSPETLISDLLMRESTENVPRTRLEDPAMW 97 C-flanking NPY S68, S69, T83 3 peptide of NPY T88, A95 P01303 29 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 64 Neuropeptide Y NPY S31, 2 (Ambiguous Y55 or T60 P01298 30 APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY 65 Pancreatic NPY T61 1 hormone P01298 69 HKEDTLAFSEWGSPHAAVPR 88 Pancreatic NPY icosapeptide P10082 29 YPIKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 64 Peptide YY NPY T60 1 P10082 31 IKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 64 Peptide YY(3- NPY T60 1 36) P80303 25 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVIDVL 106 Nesfatin-1 Nucleobindin T32, I37, S42, 8 ETDKHFREKLQKADIEEIKSGRLSKELDLVSHHVRTKLDE T50, Y53, Y54, L T67, S96 Q02818 27 VPLERGAPNKEETPATESPDTGLYYHRYLQEVIDVLETDG 461 Nucleobindin-1 Nucleobindin P34, T39, T42, 22 HFREKLQAANAEDIKSGKLSRELDFVSHHVRTKLDELKR S44, T47, Y50, QEVSRLRMLLKAKMDAEQDPNVQVDHLNLLKQFEHLDP S93, T148, QNQHTFEARDLELLIQTATRDLAQYDAAHHEEFKRYEML T162, S224, KEHERRRYLESLGEEQRKEAERKLEEQQRRHREHPKVNV S320, S321, PGSQAQLKEVWEELDGLDPNRFNPKTFFILHDINSDGVLD T335, H339, EQELEALFTKELEKVYDPKNEEDDMREMEEERLRMREHV S369, T372, MKNVDTNQDRLVTLEEFLASTQRKEFGDTGEGWETVEM S378, Q407, HPAYFEEELRRFEEELAAREAELNAKAQRLSQETEALGRS A414, T426, QGRLEAQKRELQQAVLHMEQRKQQQQQQQGHKAPAAH L452 PEGQLKFHPDTDDVPVPAPAGDQKEVDTSEKKLLERLPE x459(deleterious VEVPQHL mutation) P80303 25 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVIDVL 420 Nucleobindin-2 Nucleobindin T32, 16 ETDKHFREKLQKADIEEIKSGRLSKELDLVSHHVRTKLDE I37, LKRQEVGRLRMLIKAKLDSLQDIGMDHQALLKQFDHLNH S42, T50, Y53, LNPDKFESTDLDMLIKAATSDLEHYDKTRHEEFKKYEMM Y54, T67, S96, KEHERREYLKTLNEEKRKEEESKFEEMKKKHENHPKVNH S152, T163, PGSKDQLKEVWEETDGLDPNDFDPKTFFKLHDVNSDGFL T172, A355, DEQELEALFTKELEKVYDPKNEEDDMVEMEEERLRMRE Y389, P406, HVMSEVDTNKDRLVTLEEFLKATEKKEFLEPDSWETLDQ S408, L414 QQFFTEEELKEYENIIALQENELKKKADELQKQKEELQRQ HDQLEAQKLEYHQVIQQMEQKKLQQGIPPSGPAGELKFE PHI P01213 175 YGGFLRKYPK 184 Alpha- Opioid neoendorphin P01213 175 YGGFLRKYP 183 Beta- Opioid neoendorphin P01213 207 YGGFLRRIRPKLKWDNQKRYGGFLRRQFKVVT 238 Big dynorphin Opioid P01213 207 YGGFLRRIRPKLK 219 Dynorphin A(1- Opioid 13) (By similarity) P01213 207 YGGFLRRIRPKLKWDNQ 223 Dynorphin A(1- Opioid 17) P01213 207 YGGFLRRI 214 Dynorphin A(1- Opioid 8) (By similarity) P01213 175 YGGFL 179 Leu-enkephalin Opioid P01213 226 YGGFLRRQFKVVTRSQEDPNAYSGELFDA 254 Leumorphin Opioid P01210 100 YGGFM 104 Met-enkephalin Opioid P01210 186 YGGFMRGL 193 Met-enkephalin- Opioid Arg-Gly-Leu P01210 261 YGGFMRF 267 Met-enkephalin- Opioid Arg-Phe Q13519 98 MPRVRSLFQEQEEPEPGMEEAGEMEQKQLQ 127 Neuropeptide 1 Opioid P111 1 (Probable) Q13519 149 FSEFMRQYLVLSMQSSQ 165 Neuropeptide 2 Opioid (Probable) Q13519 130 FGGFTGARKSARKLANQ 146 Nociceptin Opioid P01210 143 DAEEDDSLANSSDLLKELLETGDNRERSHHQDGSDNEEE 183 PENK(143-183) Opioid VS (By similarity) P01210 237 FAEALPSDEEGESYSKEVPEME 258 PENK(237-258) Opioid (By similarity) P01213 226 YGGFLRRQFKVVT 238 Rimorphin Opioid P01210 114 MDELYPMEPEEEANGSEILA 133 rimorphin Opioid P01210 25 ECSQDCATCSYRLVRPADINFLACVMECEGKLPSLKIWET 97 Synenkephalin Opioid CKELLQLSKPELPQDGTSTLRENSKPEESHLLA O43612 34 QPLPDCCRQKTCSCRLYELLHGAGNHAAGILTL 66 Orexin-A Orexin O43612 70 RSGPPGLQGRLQRLLQASGNHAAGILTM 97 Orexin-B Orexin P12272 143 TRSAWLDSGVTGSGLEGDHLSDTSTTSLELDSR 175 Osteostatin Parathyroid hormone P12272 37 AVSEHQLLHDKGKSIQDLRRREFLHHLIAEIHTAEIRATSE 177 Parathyroid Parathyroid T75 1 VSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKVETYKE hormone-related hormone QPLKTPGKKKKGKPGKRKEQEKKKRRTRSAWLDSGVTG protein SGLEGDHLSDTSTTSLELDSRRH P12272 37 AVSEHQLLHDKGKSIQDLRRREFLHHLIAEIHTAEI 72 PTHrP[1-36] Parathyroid hormone P12272 74 ATSEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKVET 130 PTHrP[38-94[ Parathyroid T75 1 YKEQPLKTPGKKKKGKP hormone Q96A98 62 SLALADDAAFRERARLLAALERRHWLNSYMHKLLVLDA 100 Tuberoinfundibular Parathyroid P peptide of hormone 39residues P01189 237 YGGFMTSEKSQTPLVTLEKNAIIKNAYKKGE 267 Beta-endorphin POMC S246, T248, 3 T252 P01189 138 SYSMEHFRWGKPVGKKRRPVKVYPNGAEDESAEAFPLEF 176 Corticotropin POMC P01189 156 PVKVYPNGAEDESAEAFPLEF 176 Corticotropin- POMC like intermediary peptide P01189 179 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAAEKK 267 Lipotropin beta POMC S246, T248, 3 DEGPYRMEHFRWGSPPKDKRYGGFMTSEKSQTPLVTLFK T252 NAIIKNAYKKGE P01189 179 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAAEKK 234 Lipotropin POMC DEGPYRMEHFRWGSPPKD gamma P01189 138 SYSMEHFRWGKPV 150 Melanotropin POMC alpha P01189 217 DEGPYRMEHFRWGSPPKD 234 Melanotropin POMC beta P01189 77 YVMGHFRWDRF 87 Melanotropin POMC gamma P01189 27 WCLESSQCQDLTTESNLLECIRACKPDLSAETPMFPGNGD 102 NPP POMC T71 1 EQPLTENPRKYVMGHFRWDRFGRRNSSSSGSSGAGQ P01189 105 EDVSAGEDCGPLPEGGPEPRSDGAKPGPRE 134 Potential peptide POMC Q9UHG2 245 LETPAPQVPARRLLPP 260 Big LEN (By ProSAAS T247, P248, 2 similarity) A249 Q9UHG2 221 AADHDVGSELPPEGVLGALLRVKRLETPAPQVPARRLLPP 260 Big PEN-LEN ProSAAS T247, P248, 2 (By similarity) A249 Q9UHG2 34 ARPVKEPRGLSAASPPLAETGAPRRF 59 Big SAAS (By ProSAAS G42, S47, T53, 5 similarity) G54, A55 Q9UHG2 34 ARPVKEP 40 KEP (By ProSAAS similarity) Q9UHG2 245 LETPAPQVPA 254 Little LEN (By ProSAAS T247, P248, 2 similarity) A249 Q9UHG2 42 GLSAASPPLAETGAPRRF 59 Little SAAS (By ProSAAS G42, S47, T53, 5 similarity) G54, A55 Q9UHG2 221 AADHDVGSELPPEGVLGALLRV 242 PEN (By ProSAAS similarity) Q9UHG2 34 ARPVKEPRGLSAASPPLAETGAPRRFRRSVPRGEAAGAVQ 260 ProSAAS ProSAAS G42, S47, T53, 13 ELARALAHLLEAERQERARAEAQEAEDQQARVLAQLLR G54, A55, A115, VWGAPRNSDPALGLDDDPDAPAAQLARALLRARLDPAAL N118, S119, AAQLVPAPVPAAALRPRPPVYDDGPAGPDAEEAGDETPD A176, S206, VDPELLRYLLGRILAGSADSEGVAAPRRLRRAADHDVGSE A207, T247, LPPEGVLGALLRVKRLETPAPQVPARRLLPP P248, A249 Q9HD89 19 KTLCSMEEAINERIQEVAGSLIFRAISSIGLECQSVTSRGDL 108 Resistin Resistin/FIZZ ATCPRGFAVTGCTCGSACGSWDVRAETTCHCQCAGMDW TGARCCRVQP Q9BQ08 24 QCSLDSVMDKKIKDVLNSLEYSPSPISKKLSCASVKSQGR 111 Resistin-like beta Resistin/FIZZ PSSCPAGMAVTGCACGYGCGSWDVQLETTCHCQCSVVD WTTARCCHLT P83859 91 QDEGSEATGFLPAAGEKTSGPLGNLAEELNGYSRKKGGF 133 QRF-amide RFamide SFRF neuropeptide P06850 154 SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII 194 Corticoliberin Sauvagine/ corticotropin- releasing factor/urotensin I P55089 83 DNPSLSIDLTFHLLRTLLELARTQSQRERAEQNRIIFDSV 122 Urocortin Sauvagine/ corticotropin- releasing factor/urotensin I Q96RP3 72 IVLSLDVPIGLLQILLEQARARAAREQATTNARILARVGHC 112 Urocortin-2 Sauvagine/ corticotropin- releasing factor/urotensin I Q969E3 120 FTLSLDVPTNIMNLLFNIAKAKNLRAQAAANAHLMAQI 157 Urocortin-3 Sauvagine/ corticotropin- releasing factor/urotensin I P01019 34 DRVY 37 Angiotensin 1-4 Serpin P01019 34 DRVYI 38 Angiotensin 1-5 Serpin P01019 34 DRVYIHP 40 Angiotensin 1-7 Serpin P01019 34 DRVYIHPFH 42 Angiotensin 1-9 Serpin P01019 34 DRVYIHPFHL 43 Angiotensin-1 Serpin P01019 34 DRVYIHPF 41 Angiotensin-2 Serpin P01019 35 RVYIHPF 41 Angiotensin-3 Serpin P01019 36 VYIHPF 41 Angiotensin-4 Serpin P01019 34 DRVYIHPFHLVIHNESTCEQLAKANAGKPKDPTFIPAPIQA 485 Angiotensinogen Serpin T440 or S442 or 1 KTSPVDEKALQDQLVLVAAKLDFEDKLRAAMVGMLANF T443 or T455 LGFRIYGMHSELWGVVHGATVLSPTAVFGTLASLYLGAL (Ambiguous) DHTADRLQAILGVPWKDKNCTSRLDAHKVLSALQAVQG LLVAQGRADSQAQLLLSTVVGVFTAPGLHLKQPFVQGLA LYTPVVLPRSLDFTELDVAAEKIDRFMQAVTGWKTGCSL MGASVDSTLAFNTYVHFQGKMKGFSLLAEPQEFWVDNS TSVSVPMLSGMGTFQHWSDIQDNFSVTQVPFTESACLLLI QPHYASDLDKVEGLTFQQNSLNVVMKKLSPRTIHLTMPQL VLQGSYDLQDLLAQAELPAILHFELNLQKLSNDRIRVGEV LNSIFFELEADEREPTESTQQLNKPEVLEVTLNRPFLFAVY DQSATALHFLGRVANPLSTA P08185 23 MDPNAAYVNMSNHHRGLASANVDFAFSLYKHLVALSPK 405 Corticosteroid- Serpin KNIFISPVSISMALAMLSLGTCGHTRAQLLQGLGFNLTERS binding globulin ETEIHQGFQHLHQLFAKSDTSLEMTMGNALFLDGSLELLE SFSADIKHYYESEVLAMNFQDWATASRQINSYVKNKTQG KIVDLFSGLDSPAILVLVNYIFFKGTWTQPFDLASTREENF YVDETTVVKVPMMLQSSTISYLHDSELPCQLVQMNYVGN GTVFFILPDKGKMNTVIAALSRDTINRWSAGLTSSQVDLY IPKVTISGVYDLGDVLEEMGIADLFTNQANFSRITQDAQL KSSKVVHKAVLQLNEEGVDTAGSTGVTLNLTSKPIILRFN QPFIIMIFDHFTWSSLFLARVMNPV Q86U17 20 QPLLAHGDKSLQGPQPPRHQLSEPAPAYHRITPTITNFALR 422 Serpin A11 Serpin LYKELAADAPGNIFFSPVSISTTLALLSLGAQANTSALILEG LGFNLTETPEADIHQGFRSLLHTLALPSPKLELKVGNSLFL DKRLKPRQHYLDSIKELYGAFAFSANFTDSVTTGRQINDY LRRQTYGQVVDCLPEFSQDTFMVLANYIFFKAKWKHPFS RYQTQKQESFFVDERTSLQVPMMHQKEMHRFLYDQDLA CTVLQIEYRGNALALLVLPDPGKMKQVEAALQPQTLRK WGQLLLPSLLDLHLPRFSISGTYNLEDILPQIGLTNILNLEA DFSGVTGQLNKTISKVSHKAMVDMSEKGFLAGAASGLLS QPPSLNTMSDPHAHFNRPFLLLLWEVTTQSLLFLGKVVNP VAG Q8IW75 21 LKPSFSPRNYKALSEVQGWKQRMAAKELARQNMDLGFK 414 Serpin A12 Serpin LLKKLAFYNPGRNIFLSPLSISTAFSMLCLGAQDSTLDEIK QGFNFRKMPEKDLHEGFHYIIHELTQKTQDLKLSIGNTLFI DQRLQPQRKFLEDAKNFYSAETILTNFQNLEMAQKQINDF ISQKTHGKINNLIENIDPGTVMLLANYIFFRARWKHEFDPN VTKEEDFFLEKNSSVKVPMMFRSGIYQVGYDDKLSCTILE IPYQKNITAIFILPDEGKLKHLEKGLQVDTFSRWKTLLSRR VVDVSVPRLHMTGTFDLKKTLSYIGVSKIFLEHGDLTKIA PHRSLKVGEAVHKAELKMDERGFEGAAGTGAQTLPMET PLVVKIDKPYLLLIYSEKIPSVLFLGKIVNPIGK O75830 19 SRCSAQKNTEFAVDLYQEVSLSHKDNIIFSPLGITLVLEMV 405 Serpin 12 Serpin QLGAKGKAQQQIRQTLKQQETSAGEEFFVLKSFFSAISEK KQEFTFNLANALYLQEGFTVKEQYLHGNKEFFQSAIKLV DEQDAKACAEMISTWVERKTDGKIKDMFSGEEFGPLTRL VLVNAIYFKGDWKQKFRKEDTQLINFTKKNGSTVKIPMM KALLRTKYGYFSESSLNYQVLELSYKGDEFSLIIILPAEGM DIEEVEKLITAQQILKWLSEMQEEEVEISLPREKVEQKVDF KDVLYSLNITEIFSGGCDLSGITDSSEVYVSQVTQKVFFEIN EDGSEAATSTGIHIPVIMSLAQSQFIANHPFLFIMKHNPTES ILFMGRVTNPDTQEIKGRDLDSL O00230 89 DRMPCRNFFVVICITSSCK 105 Cortistatin-17 Somastostatin O00230 77 QEGAPPQQSARRDRMPCRNFFWKTFSSCK 105 Cortistatin-29 Somastostatin (Potential) P61278 103 AGCKNFFWKTFTSC 116 Somatostatin-14 Somastostatin P61278 89 SANSNPAMAPRERKAGCKNFFWKTFTSC 116 Somatostatin-28 Somastostatin S89 or S92 1 (Ambiguous) P01236 29 LPICPGGAARCQVTLRDLFDRAVVLSHYIHNLSSEMFSEF 227 Prolactin Somatotropin/ DKRYTHGRGFITKAINSCHTSSLATPEDKEQAQQMNQKD prolactin FLSLIVSILRSWNEPLYHLVTEVRGMQEAPEAILSKAVEIE EQTKRLLEGMELIVSQVHPETKENEIYPVWSGLPSLQMAD EESRLSAYYNLLHCLRRDSHKIDNYLKLLKCRIIHNNNC P20366 111 ALNSVAYERSAMQNYE 125 C-terminal- Tachykinin A111, S114 2 flanking peptide P20366 98 HKTDSFVGLM 107 Neurokinin A Tachykinin Q9UHF0 81 DMHDFFVGLM 90 Neurokinin-B Tachykinin P20366 72 DADSSIEKQVALLKALYGHGQISHKRHKTDSFVGLM 107 Neuropeptide K Tachykinin P20366 58 RPKPQQFFGLM 68 Substance P Tachykinin P20396 25 QPEAAQQEAVTAAEHPGLDDFLRQVERLLFLRENIQRLQ 242 Pro-thyrotropin- TRH 174 1 GDQGEHSASQIFQSDWLSKRQHPGKREEEEEEGVEEEEEE releasing EGGAVGPHKRQHPGRREDEASWSVDVTQHKRQHPGRRS hormone PWLAYAVPKRQHPGRRLADPKAQRSWEEEEEEEEREEDL MPEKRQHPGKRALGGPCGPQGAYGQAGLLLGLLDDLSRS QGAEEKRQHPGRRAAWVREPLEE P20396 84 QHP 86 Thyrotropin- TRH releasing hormone O95399 114 ETPDCFWKYCV 124 Urotensin-2 Urotensin-2 Q76510 112 ACFVVKYCV 119 Urotensin-2B Urotensin-2 P01185 20 CYFQNCPRG 28 Arg-vasopressin Vasopressin/ oxytocin P01185 126 ASDRSNATQLDGPAGALLLRLVQLAGAPEPFEPAQPDAY 164 Copeptin Vasopressin/ A152 1 oxytocin P01178 32 AAPDLDVRKCLPCGPGGKGRCFGPNICCAEELGCFVGTA 125 Neurophysin 1 Vasopressin/ EALRCQEENYLPSPCQSGQKACGSGGRCAVLGLCCSPDG oxytocin CHADPACDAEATFSQR P01185 32 AMSDLELRQCLPCGPGGKGRCFGPSICCADELGCFVGTAE 124 Neurophysin 2 Vasopressin/ ALRCQEENYLPSPCQSGQKACGSGGRCAAFGVCCNDESC oxytocin VTEPECREGFHRRA P01178 20 CYIQNCPLG 28 Oxytocin Vasopressin/ oxytocin O15240 554 TLQPPSALRRRHYHHALPPSRHYP 577 Antimicrobial VGF peptide VGF[554-577] O15240 281 RPESALLGGSEAGERLLQQGLAQVEA 306 Neuroendocrine VGF regulatory peptide-1 O15240 310 QAEATRQAAAQEERLADLASDLLLQYLLQGGARQRGLG 347 Neuroendocrine VGF regulatory peptide-2 O15240 25 APPGRPEAQPPPLSSEHKEPVAGDAVPGPKDGSAPEVRGA 615 Neurosecretory VGF P25, S36, P226, 5 RNSEPQDEGELFQGVDPRALAAVLLQALDRPASPPAPSGS protein VGF S229, T501 QQGPEEEAAEALLTETVRSQTHSLPAPESPEPAAPPRPQTP ENGPEASDPSEELEALASLLQELRDFSPSSAKRQQETAAA ETETRTHTLTRVNLESPGPERVWRASWGEFQARVPERAP LPPPAPSQFQARMPDSGPLPETHKFGEGVSSPKTHLGEAL APLSKAYQGVAAPFPKARRPESALLGGSEAGERLLQQGL AQVEAGRRQAEATRQAAAQEERLADLASDLLLQYLLQG GARQRGLGGRGLQEAAEERESAREEEEAEQERRGGEERV GEEDEEAAEAEAEAEEAERARQNALLFAEEEDGEAGAED KRSQEETPGHRRKEAEGTEEGGEEEDDEEMDPQTIDSLIE LSTKLHLPADDVVSIIEEVEEKRKRKKNAPPEPVPPPRAAP APTHVRSPQPPPPAPAPARDELPDWNEVLPPWDREEDEV YPPGPYHPFPNYIRPRTLQPPSALRRRHYHHALPPSRHYPG REAQARRAQEEAEAEERRLQEQEELENYIEHVLLRRP
Specific embodiments of the invention are disclosed below with indication of the respective SEQ ID NO.
TABLE-US-00007 TABLE 6A *Site position (Bold; identified SEQ Peptide or conserved in human, Italic: No. ID Entry_ Peptide Peptide Peptide Hormone identified in non-human and not of NO Human Start Peptide_Sequence end Name Family conserved sites 147 P01160 1 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Atrial Natriuretic S19N, S25 2 natriuretic peptide factor
TABLE-US-00008 TABLE 6B *Site position (Bold; identified SEQ Peptide or conserved in human, Italic: No. ID Entry_ Peptide Peptide Peptide Hormone identified in non-human and not of NO Human Start Peptide_Sequence end Name Family conserved sites 72 P22466 1 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 30 Galanin Galanin T3, S23 2 95 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 29 Glucagon Glucagon T7 1 97 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 31 Glucago-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1 (7-36) 106 P09683 1 HSDGTFTSELSRLREGARLQRLLQGLV 27 Secretin Glucagon T7, S8, S11 3 108 P01282 1 HSDAVFTDNYTRLRKQMAVKKYLNSILN 28 Vasoactive Glucagon T7 1 intestinal peptide 147 P01160 1 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Atrial Natriuretic S19, S25 2 natriuretic peptide factor 185 P01303 1 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 36 Neuropeptide Y NPY S3, T32 2 188 P10082 1 YPIKPAPREDASPEELNRYYASLRHYLNLVTRQRY 36 Peptide YY NPY T32 1
TABLE-US-00009 TABLE 6C SEQ Peptide No. ID Entry_ Peptide Peptide Peptide Hormone *Site position (identified of NO Human Start Peptide_Sequence end Name Family or conserved in humans) sites 92 P09681 1 YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDW 42 Gastric Glucagon S8 1 KHNITQ inhibitory polypeptide 95 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 29 Glucagon Glucagon T7 1 97 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 31 Glucago-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1 (7-36) 99 P01275 1 HADGSFSDEMNTILDNLAARDFINWLIQTKITD 33 Glucagon-like Glucagon S7 1 peptide 2 (By similarity) 100 P01282 1 HADGVFTSDFSKLLGQLSAKKYLESLM 27 Intestinal Glucagon T7, S8 2 peptide PHM-27 106 P09683 1 HSDGTFTSELSRLREGARLQRLLQGLV 27 Secretin Glucagon T7, S8, S11 3 107 P01286 1 YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQ 44 Somatoliberin Blucagon T7 1 ERGARARL 108 P01282 1 HSDAVFTDNYTRLRKQMAVKKYLNSILN 28 Vasoactive Glucagon T7 1 intestinal peptide 21 P01258 1 CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP 32 Calcitonin Calcitonin T6, T11. T13, T21, T25 5 22 P06881 1 ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSK 37 Calcitonin Calcitonin T9, S17 2 AF gene- related peptide 1 23 P10092 1 ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSK 37 Calcitonin Calcitonin T9, S17, S25, T30, S34 5 AG gene- related peptide 2 24 P10997 1 KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSN 37 Islet amyloid Calcitonin T6, T9 3 TY polypeptide 6 P35318 1 YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTD 52 Adrenomedullin Adrenomedullin T22, Y31, T34 3 KDKDNVAPRSKISPQGY 7 Q7Z4H4 1 TQAQLLRVGCVLGTCQVQNLSHRLWQLMGPAGRQDS 47 Adrenomedullin- Adrenomedullin T14 1 APVDPSSPHSY 2 (By similarity) 116 P01308 1 FVNQHLCGSHLVEALYLVCGERGFFYTPKT 30 Insulin B Insulin T27 (Ambiguous) 1 chain 117 P05019 1 GPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSS 70 Inslulin-like Insulin T29 1 RAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA growth factor I 118 P01344 1 AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVS 67 Inslulin-like Insulin S29, S50, T62 2 RRSRGIVEECCFRSCDLALLETYCATPAKSE growth factor II 72 P22466 1 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 30 Galanin Galanin T3, S23 2 74 Q9UBC7 1 APAHRGRGGWTLNSAGYLLGPVLHLPQMGDQDGKRE 60 Galanin-like Galanin T11 1 TALEILDLWKAIDGLPYSHPPQPS peptide 185 P01303 1 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 36 Neuropeptide Y NPY S3, T32 2 186 P01298 1 APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY 36 Pancreatic NPY T32 1 hormone 188 P10082 1 YPIKPAPREDASPEELNRYYASLRHYLNLVTRQRY 36 Peptide YY NPY T32 1 147 P01160 1 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Atrial Natriuretic S19, S25 2 natriuretic peptide factor 163 P16860 1 SPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH 32 Brain Natriuretic S22 1 natriuretic peptide 167 P23582 1 DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKG 53 CNP-53 Natriuretic S47 1 CFGLKLDRIGSMSGLGC peptide
TABLE-US-00010 TABLE 6D *Site position (Bold: identified or conserved SEQ Pep- Pep- Peptide in human, Italic; No. ID Entry_ tide tide Peptide Hormone identified in non-human of NO. Human Start Peptide_Sequence end Name Family and not conserved) sites 1 P05408 1 SVPHFSDEDKDPE 13 C-terminal 7B2 S1 1 peptide (By similarity) 2 P05408 1 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQ 186 Neuroendocrine 7B2 S2, T5, S10, T108, S174 5 AMNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAELTGDN protein 7B2 IPKDFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAEF SREFQLHQHLFDPEHDYPGLGKWNKKLLYEKMKGGE RRKRRSVNPYLQGQRLDNVVAKKSVPHFSDEDKDPE 3 P05408 1 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPAHQ 150 N-terminal 7B2 S2, T5, S10, T108 4 AMNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAELTGDN peptide (By IPKDFSEDQGYPDPPNPCPVGKTADDGCLENTPDTAEF similarity) SREFQLHQHLFDPEHDYPGLGKWNKKLLYEKMKGGE 5 P07108 1 SQAEFEKAAEEVRHLKTKPSDEEMLFIYGHYKQATVG 86 Acyl-CoA- ACBP T35 or T41 (ambiguous) 1 DINTERPGMLDFTGKAKWDAWNELKGTSKEDAMKA binding protein YINKVEELKKKYGI 6 P35318 1 YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTDKD 52 Adrenomedullin Adrenomedullin T22, Y31, T34 3 KDNVAPRSKISPQGY 7 Q7Z4H4 1 TQAQLLRVGCVLGTCQVQNLSHRLWQLMGPAGRQDS 47 Adrenomedullin-2 Adrenomedullin T14 1 APVDPSSPHSY (By similarity) 14 P07492 1 VPLPAGGGTVLTKMYPRGNHWAVGHLM 27 Gastrin-releasing Bombesin/neuromedin- P4, T9 2 peptide B/ranatensin 15 P08949 1 GNLWATGHFM 10 Neuromedin-B Bombesin/neuromedin- T6 1 B/ranatensin 16 P08949 1 APLSWDLPEPRSRASKIRVHSRGNLWATGHFM 32 Neuromedin-B-32 Bombesin/neuromedin- T28 1 B/ranatensin 21 P01258 1 CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP 32 Calcitonin Calcitonin T6, T11, T13, T21, T25 5 22 P06881 1 ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSKAF 37 Calcitonin gene- Calcitonin T9, S17 2 related peptide 1 23 P10092 1 ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSKAF 37 Calcitonin gene- Calcitonin T9, S17, S25, T30, S34 5 related peptide 2 24 P10997 1 KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY 37 Islet amyloid Calcitonin T5, T9, T30 3 polypeptide 25 P01258 1 DMSSDLERDHRPHVSMPQNAN 21 Katacalcin Calcitonin S15 1 36 P05060 1 SAEFPDFYDSEEPVSTHQEAENEKDRADQTVLTEDEKK 57 CCB peptide Chromogranin/ S15 1 ELENLAAMDLELQKIAEKF secretogranin 37 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQEC 439 Chromogranin-A Chromogranin/ S27, S30, S36, S80, E104, S108, 39 FETLRGDERILSILRHQNLLKELQDLALQGAKERAHQQ secretogranin A127, T128, A135, L136, S143 KKHSGFEDELSEVLENQSSQAELKEAVEEPSSKDVME A151, N164, T165 P167, A169, KREDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQ S170, S173, E182, L188 S189, APGEEEEEEEEATNTHPPASLPSQKYPGPQAEGDSEGLS V193, S200, T233, L236, N237, QGLVDREKGLSAEPGWQAKREEEEEEEEEAEAGEEAV P238, S241, Y244, P265, E268, PEEEGPTVVLNPHPSLGYKEIRKGESRSEALAVDGAGK P273, S282, M291, V294, T335, PGAEEAQDPEGKGEQEHSQQKEEEEEMAVVPQGLFRG A360, S380, G395 GKSGELEQEEERLSKEWEDSKRWSKMDQLAKELTAE KRLEGQEEEEDNRDSSMKLSFRARAYGFRGPGPQLRR GWRPSSREDSLEAGLPLQVRGYPEEKKEEEGSANRRPE DQELESLSAIEAELEKVAHQLQALRRG 38 P10645 1 EDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQAP 92 EA-92 Chromogranin/ A12, T13, A20, L21, S28, A36, 17 GEEEEEEEEATNTHPPASLPSQKYPGPQAEGDSEGLSQ secretogranin N49, T50, P52, A54, S55, S58, GLVDREKGLSAEPGWQA E67, L73, S74, V78, S85 39 P10645 1 EEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRR 37 ER-37 Chromogranin/ S17 or S19 (Ambiguous) 1 secretogranin 40 P10645 1 EEEEEEEEEAEAGEEAVPEEEGPTVVLNPHPSL 33 ES-43 Chromogranin/ T24, L27, N28, P29, S32 5 secretogranin 41 P05060 1 FLGEGHHRVQENQMDKARRHPQGAWKELDRNYLNY 74 GAWK peptide Chromogranin/ H7, N12, Y35, T72, A73 5 GEEGAPGKWQQQGDLQDTKENREEARFQDKQYSSHH secretogranin TAE 42 P10645 1 GYPEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQ 44 GR-44 Chromogranin/ G1 1 LQALRR secretogranin 43 P10645 1 GWRPSSREDSLEAGLPLQV 19 GV-19 Chromogranin/ S6 1 secretogranin 45 P10645 1 SEALAVDGAGKPGAEEAQDPEGKGEQEHSQQKEEEEE 48 Pancreastatin Chromogranin/ P12, E15, P20, S29, M38, V41 6 MAVVPQGLFRG secretogranin 46 P05060 1 MPVDNRNHNEGMVTRCIIEVLSNALSKSSAPPITPECR 657 Secretogranin-1 Chromogranin/ T14, S26, S28, S29, P32, T59, 40 QVLKTSRKDVKDKETTENENTKFEVRLLRDPADASEA secretogranin S73, A75, S80, A84, E89, T96, HESSSRGEAGAPGEEDIQGPTKADTEKWAEGGGHSRE S120, S124, S129, T174, N176, RADEPQWSLYPSDSQVSEEVKTRHSEKSQREDEEEEEG S186, S273, S274, S278, P287, ENYQKGERGEDSSEEKHLEEPGETQNAFLNERKQASAI S22L, M298, S300, D306, T310, KKEELVARSETHAAGHSQEKTHSREKSSQESGEETGSQ Y328, P329, G330, K376, M377, ENHPQESKGQPRSQEESEEGEEDATSEVDKRRTRPRHH H426, Y454, T491, A492, T536, HGRSRPDRSSQGGSLPSEEKGHPQEESEESNVSMASLG S557, N568, S611 EKRDHHSTHYRASEEEPEYGEEIKGYPGVQAPEDLEW ERYRGRGSEEYRAPRPQSEESWDEEDKRNYPSLELDK MAHGYGEESEEERGLEPGKGRHHRGRGGEPRAYFMS DTREEKRFLGEGHHRVQENQMDKARRHPQGAWKELD RNYLNYGEEGAPGKWQQQGDLQDTKENREEARFQDK QYSSHHTAEKRKRLGELFNPYYDPLQWKSSHFERRDN MNDNFLEGEEENELTLNEKNFFPEYNYDWWEKKPFSE DVNWGYEKRNLARVPKLDLKRQYDRVAQLDQLLHY RKKSAEFPDFYDSEEPVSTHQEAENEKDRADQTVLTE DEKKELENLAAMDLELQKIAEKFSQRG 47 P13521 1 QRNQLLQKEPDLRLENVQKFPSPEMIRALEYIENLRQQ 587 Secretogranin-2 Chromogranin/ S22, S45, A106, Y160, T161, 16 AHKEESSPDYNPYQGVSVPLQQKENGDESHLPERDSLS secretogranin S164, T167, S229, T231, S348, EEDWMRIILEALRQAENEPQSAPKENKPYALNSEKNFP T420, N459, G499, S526, S536, MDMSDDYETQQWPERKLKHMQFPPMYEENSRDNPFK P543 RTNEIVEEQYTPQSLATLESVFQELGKLTGPNNQKRER MDEEQKLYTDDEDDIYKANNIAYEDVVGGEDWNPVE EKIESQTQEEVRDSKENIEKNEQINDEMKRSGQLGIQEE DLRKESKDQLSDDVSKVIAYLKRLVNAAGSGRLQNGQ NGERATRLFEKPLDSQSIYQLIEISRNLQIPPEDLIEMLK TGEKPNGSVEPERELDLPVDLDDISEADLDHPDLFQNR MLSKSGYPKTPGRAGTEALPDGLSVEDILNLLGMESA ANQKTSYFPNPYNQEKVLPRLPYGAGRSRSNQLPKAA WIPHVENRQMAYENLNDKDQELGEYLARMLVKYPEII NSNQVKRVPGQGSSEDDLQEEEQIEQAIKEHLNQGSSQ ETDKLAPVSKRFPVGPPKNDDTPNRQYWDEDLLMKV LEYLNQEKAEKGREHIAKRAMENM 48 Q8WXD2 1 FPKPGGSQDKSLHNRELSAERPLNEQIAEAEEDKIKKT 449 Secretogranin-3 Chromogranin/ P41, T63, S103, T104, T125, 15 YPPENKPGQSNYSFVDNLNLLKAITEKEKIEKERQSIRS secretogranin A138, D192, P193, T197, S198, SPLDNKLNVEDVDSTKNRKLIDDYDSTKSGLDHKFQD T200, T212, I217, A222, S340 DPDGLHQLDGTPLTAEDIVHKIAARIYEENDRAVFDKI VSKLLNLGLITESQAHTLEDEVAEVLQKLISKEANNYE EDPNKPTSWTENQAGKIPEKVTPMAAIQDGLAKGEND ETVSNTLTLTNGLERRTKTYSEDNFEELQYFPNFYALL KSIDSEKEAKEKETLITIMKTLIDFVKMMVKYGTISPEE GVSYLENLDEMIALQTKNKLEKNATDNISKLFPAPSEK SHEETDSTKEEAAKMEKEYGSLKDSTKDDNSNPGGKT DEPKGKTEAYLEAIRKNIEWLKKHDKKGNKEDYDLSK MRDFINKQADAYVEKGILDKEEAEAIKRIYSSL 49 P13521 1 TNEIVEEQYTPQSLATLESVFQELGKLTGPNNQ 33 Secretoneurin Chromogranin/ Y9, T10, S13, T16, 4 secretogranin 51 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQEC 76 Vasostatin-1 Chromogranin/ S27, S30, S36 3 FETLRGDERILSILRHQNLLKELQDLALQGAKERAHQQ secretogranin 52 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQEC 113 Vasostatin-2 Chromogranin/ S27, S30, S36, S80, E104, 6 FETLRGDERILSILRHQNLLKELQDLALQGAKERAHQQ secretogranin S108 KKHSGFEDELSEVLENQSSQAELKEAVEEPSSKDVME 54 P10645 1 WSKMDQLAKELTAE 14 WE-14 Chromogranin/ T12 1 secretogranin 55 P01042 1 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFV 626 Kininogen-1 Cystatin T119, T133, T243, T255, E381, 15 LYRITEATKTVGSDTFYSFKYEIKEGDCPVQSGKTWQD T382, T383, S385, T389, S390, CEYKDAAKAATGECTATVGKRSSTKFSVATQTCQITP T528, S586, T592, S593, T610 AEGPVVTAQYDCLGCVHPISTQSPDLEPILRHGIQYFN NNTQHSSLFMLNEVKRAQRQVVAGLNFRITYSIVQTN CSKENFLFLTPDCKSLWNGDTGECTDNAYIDIQLRIASF SQNCDIYPGKDFVQPPTKICVGCPRDIPTNSPELEETLT HTITKLNAENNATFYFKIDNVKKARVQVVAGKKYFID FVARETTCSKESNEELTESCETKKLGQSLDCNAEVYVV PWEKKIYPTVNCQPLGMISLMKRPPGFSPFRSSRIGEIK EETTVSPPHTSMAPAQDEERDSGKEQGHTRRHDWGH EKQRKHNLGHGHKHERDQGHGHQRGHGLGHGHEQQ HGLGHGHKFKLDDDLEHQGGHVLDHGHKHKHGHGH GKHKNKGKKNGKHNGWKTEHLASSSEDSTTPSAQTQ EKTEGPTPIPSLAKPGVTVTFSDFQDSDLIATMMPPISP APIQSDDDWIPDIQIDPNGLSFNPISDFPDTTSPKCPGRP WKSVSEINPTTQMKESYYFDLTDGLS 56 P01042 1 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQFV 362 Kininogen-1 Cystatin T119, T133, T243, T255 4 LYRITEATKTVGSDTFYSFKYEIKEGDCPVQSGKTWQD heavy chain CEYKDAAKAATGECTATVGKRSSTKFSVATQTCQITP AEGPVVTAQYDCLGCVHPISTQSPDLEPILRHGIQYFN NNTQHSSLFMLNEVKRAQRQVVAGLNFRITYSIVQTN CSKENFLFLTPDCKSLWNGDTGECTDNAYIDIQLRIASF SQNCDIYPGKDFVQPPTKICVGCPRDIPTNSPELEETLT HTITKLNAENNATFYFKIDNVKKARVQVVAGKKYFID FVARETTCSKESNEELTESCETKKLGQSLDCNAEVYVV PWEKKIYPTVNCQPLGMISLMK 57 P01042 1 SSRIGEIKEETTVSPPHTSMAPAQDEERDSGKEQGHTR 255 Kininogen-1 light Cystatin E10, T11, T12, S14, T18, S19, 11 RHDWGHEKQRKHNLGHGHKHERDQGHGHQRGHGLG chain T157, S215, T221, S222, T239 HGHEQQHGLGHGHKFKLDDDLEHQGGHVLDHGHKH KHGHGHGKHKNKGKKNGKHNGWKTEHLASSSEDSTT PSAQTQEKTEGPTPIPSLAKPGVTVTFSDFQDSDLIATM MPPISPAPIQSDDDWIPDIQIDPNGLSFNPISDFPDTT SPKCPGRPWKSVSEINPTTQMKESYYFDLTDGLS 72 P22466 1 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 30 Galanin Galanin T3, S23 2 73 P22466 1 ELRPEDDMKPGSFDRSIPENNIMRTIIEFLSFLHLKEA 59 Galanin message- Galanin A49 1 GALDRLLDLPAAASSEDIERS associated peptide 74 Q9UBC7 1 APAHRGRGGWTLNSAGYLLGPVLHLPQMGDQDGKRE 60 Galanin-like Galanin T11 1 TALEILDLWKAIDGLPYSHPPQPS peptide 76 P06307 1 QPVPPADPAGSGLQRAEEAPRRQLRVSQRTDGESRAH 95 Cholecystokinin Gastrin/ A8, T30, N65 3 LGALLARYIQQARKAPSGRMSIVKNLQNLDPSHRISDR cholecystokinin DYMGWMDFGRRSAEEYEYPS 79 P06307 1 IVKNLQNLDPSHRISDRDYMGWMDF 25 Cholecystokinin- Gastrin/ N7 1 25 (By cholecystokinin similarity) 80 P06307 1 KAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 33 Cholecystokinin- Gastrin/ N15 1 33 cholecystokinin 81 P06307 1 YIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYMGWM 39 Cholecystokinin- Gastrin/ N21 1 DF 39 cholecystokinin 83 P06307 1 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKN 58 Cholecystokinin- Gastrin/ T5, N40 2 LQNLDPSHRISDRDYMGWMDF 58 cholecystokinin 84 P06307 1 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIVKN 49 Cholecystokinin- Gastrin/ T5, N40 2 LQNLDPSHRISD 58 cholecystokinin desnonopeptide (By similarity) 91 P01350 1 SWKPRSQQPDAPLGTGANRDLELPWLEQQGPASHHRR 71 Gastrin-71 Gastrin/ L12 or G13 (Ambiguous) 1 QLGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF cholecystokinin 92 P09681 1 YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDW 42 Gastric Glucagon S8 1 KHNITQ inhibitory polypeptide 93 P01275 1 RSLQDTEEKSRSFSASQADPLSDPDQMNEDKRHSQGTF 69 Glicentin (By Glucagon S12, T39 2 TSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA similarity) 94 P01275 1 RSLQDTEEKSRSFSASQADPLSDPDQMNED 30 Glicentin-related Glucagon S12 1 polypeptide (By similarity) 95 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 29 Glucagon Glucagon T7 1 96 P01275 1 HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 37 Glucagon-like Glucagon T11 or T13 or S14 (Ambiguous) 1 peptide 1 97 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 31 Glucagon-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1(7-36) 98 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 30 Glucagon-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1(7-37) 99 P01275 1 HADGSFSDEMNTILDNLAARDFINWLIQTKITD 33 Glucagon-like Glucagon S7 1 peptide 2 (By similarity) 100 P01282 1 HADGVFTSDFSKLLGQLSAKKYLESLM 27 Intestinal Glucagon T7, S8 2 peptide PHM-27 101 P01282 1 HADGVFTSDFSKLLGQLSAKKYLESLMGKRVSSNISED 42 Intestinal Glucagon T7, S8 2 PVPV peptide PHV-42 102 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNI 3 7 Oxyntomodulin Glucagon T7 1 A (By similarity) 106 P09683 1 HSDGTFTSELSRLREGARLQRLLQGLV 27 Secretin Glucagon T7, S8, S11 3 107 P01286 1 YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQE 44 Somatoliberin Glucagon T7 1 RGARARL 108 P01282 1 HSDAVFTDNYTRLRKQMAVKKYLNSILN 28 Vasoactive Glucagon T7 1 intestinal peptide 109 P01148 1 DAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLRDL 56 GnRH-associated GnRH T28 1 KGALESLIEEETGQKKI peptide 1 113 P01148 1 QHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQR 69 Progonadoliberin- GnRH T41 1 FECTTHQPRSPLRDLKGALESLIEEETGQKKI 1 116 P01308 1 FVNQHLCGSHLVEALYLVCGERGFFYTPKT 30 Insulin B chain Insulin T27 (Ambiguous) 1 117 P05019 1 GPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSRR 70 Insulin-like Insulin T29 1 APQTGIVDECCFRSCDLRRLEMYCATPLKPAKSA growth factor I 118 P01344 1 AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRR 67 Insulin-like Insulin S50, T62 2 SRGIVEECCFRSCDLALLETYCATPAKSE growth factor II 119 P01344 1 YRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRS 66 Insulin-like Insulin S49, T61 2 RGIVEECCFRSCDLALLETYCATPAKSE growth factor II Ala-25 Del 120 P01344 1 DVSTPPTVLPDNFPRYPVGKFFQYDTWKQSTQRL 34 Preptin Insulin S3, T4, T7 3 130 Q15726 1 EPLEKVASVGNSRPTGQQLESLGLLAPGEQSLPCTERK 119 Metastasis- KISS1 A69 1 PAATARLSRRGTSLSPPPESSGSPQQPGLSAPHSRQIPA suppressor KiSS-1 PQGAVLVQREKDLPNYNWNSFGLRFGKREAAPGNHGR SAGRG 131 Q15726 1 GTSLSPPPESSGSPQQPGLSAPHSRQIPAPQGAVLVQRE 54 Metastin KISS1 A21 1 KDLPNYNWNSFGLRF 135 P20382 1 GSVAFPAENGVQNTESTQE 19 Neuropeptide- Melanin- T14, T16, T17 3 glycine-glutamic concentrating acid (Potential) hormone 136 P20382 1 ILLSASKSIRNLDDDMVFNTFRLGKGFQKEDTAEKSVI 144 Pro-MCH Melanin- S36, S41, N72, T102, T104, 6 APSLEQYKNDESSFMNEEENKVSKNTGSKHNFLNHGL concentrating T105 PLNLAIKPYLALKGSVAFPAENGVQNTESTQEKREIGD hormone EENSAKFPIGRRDFDMLRCMLGRVYRPCWQV 143 Q15848 1 ETTTQGPGVLLPLPKGACTGWMAGIPGHPGHNGAPGR 226 Adiponectin NA T4 1 DGRDGTPGEKGEKGDPGLIGPKGDIGETGVPGAEGPR GFPGIQGRKGEPGEGAYVYRSAFSVGLETYVTIPNMPI RFTKIFYNQQNHYDGSTGKFHCNIPGLYYFAYHITVYM KDVKVSLFKKDKAMLFTYDQYQENNVDQASGSVLLH LEVGDQVWLQVYGEGERNGLYADNDNDSTFTGFLLY HDTN 144 O00253 1 AQMGLAPMEGIRRPDQALLPELPGLGLRAPLKKTTAE 111 Agouti-related NA T35 1 QAEEDLLQEAQALAEVLDLQDREPRSSRRCVRLHESC protein LGQQVPCCDPCATCYCRFFNAFCYCRKLGTAMNPCSR T 147 P01160 1 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Atrial Natriuretic peptide S19, S25 2 natriuretic factor 163 P16860 1 SPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH 32 Brain natriuretic Natriuretic peptide S22 1 peptide 32 164 P01160 1 NPMYNAVSNADLMDFKNLLDHLEEKMPLED 30 Cardiodilatin- Natriuretic peptide Y4, A6, S8 (Ambiguous) 1 related peptide 167 P23582 1 DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKG 53 CNP-53 Natriuretic peptide S47 1 CFGLKLDRIGSMSGLGC 168 P16860 1 HPLGSPGSASDLETSGLQEQRNHLQGKLSELQVEQTSL 108 Natriuretic Natriuretic peptide T48, E59 2 EPLQESPRPTGVWKSREVATEGIRGHRKMVLYTLRAP peptides B RSPKMVQGSGCFGRKMDRISSSSGLGCKVLRRH 170 O95156 1 KEVVHATEGLDWEDKDAPGTLVGNVVHSRIISPLRLF 242 Neurexophilin-2 Neurexophilin T7 1 VKQSPVPKPGPMAYADSMENFWDWLANITEIQEPLAR TKRRPIVKTGKFKKMFGWGDFHSNIKTVKLNLLITGKI VDHGNGTFSVYFRHNSTGLGNVSVSLVPPSKVVEFEVS PQSTLETKESKSFNCRIEYEKTDRAKKTALCNFDPSKIC YQEQTQSHVSWLCSKPFKVICIYIAFYSVDYKLVQKVC PDYNYHSETPYLSSG 184 P01303 1 SSPETLISDLLMRESTENVPRTRLEDPAMW 30 C-flanking NPY S1, S2, T16, T21, A28 3 peptide of NPY 185 P01303 1 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 36 Neuropeptide Y NPY S3, T32 2 186 P01298 1 APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY 36 Pancreatic NPY T32 1 hormone 188 P10082 1 YPIKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 36 Peptide YY NPY T32 1 189 P10082 1 IKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 34 Peptide YY(3-36) NPY T30 1 190 P80303 1 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVID 82 Nesfatin-1 Nucleobindin T8, I13, S18, T26, Y29, Y30, 8 VLETDKHFREKLQKADIEEIKSGRLSKELDLVSHHVRT T43, S72 KLDEL 191 Q02818 1 VPLERGAPNKEETPATESPDTGLYYHRYLQEVIDVLET 435 Nucleobindin-1 Nucleobindin P8, T13, T16, S18, T21, Y24, 22 DGHFREKLQAANAEDIKSGKLSRELDFVSHHVRTKLD S67, T122, T136, S198, S294, ELKRQEVSRLRMLLKAKMDAEQDPNVQVDHLNLLKQ S295, T309, H313, S343, T346, FEHLDPQNQHTFEARDLELLIQTATRDLAQYDAAHHE S352, Q381, A388, T400, L1126, EFKRYEMLKEHERRRYLESLGEEQRKEAERKLEEQQR x433 (deleterious mutation) RHREHPKVNVPGSQAQLKEVWEELDGLDPNRFNPKTF FILHDINSDGVLDEQELEALFTKELEKVYDPKNEEDDM REMEEERLRMREHVMKNVDTNQDRLVTLEEFLASTQ RKEFGDTGEGWETVEMHPAYTEEELRRFEEELAAREA ELNAKAQRLSQETEALGRSQGRLEAQKRELQQAVLH MEQRKQQQQQQQGHKAPAAHPEGQLKFHPDTDDVPV PAPAGDQKEVDTSEKKLLERLPEVEVPQHL 192 P80303 1 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQVID 396 Nucleobindin-2 Nucleobindin T8, I13, S18, T26, Y29, Y30, 16 VLETDKHFREKLQKADIEEIKSGRLSKELDLVSHHVRT T43, S72, S128, T139, T148, KLDELKRQEVGRLRMLIKAKLDSLQDIGMDHQALLKQ A331, Y365, P382, S384, L390 FDHLNHLNPDKFESTDLDMLIKAATSDLEHYDKTRHE EFKKYEMMKEHERREYLKTLNEEKRKEEESKFEEMKK KHENHPKVNHPGSKDQLKEVWEETDGLDPNDFDPKTF FKLHDVNSDGFLDEQELEALFTKELEKVYDPKNEEDD MVEMEEERLRMREHVMSEVDTNKDRLVTLEEFLKAT EKKEFLEPDSWETLDQQQFFTEEELKEYENIIALQENEL KKKADELQKQKEELQRQHDQLEAQKLEYHQVIQQME QKKLQQGIPPSGPAGELKFEPHI 204 Q13519 1 MPRVRSLFQEQEEPEPGMEEAGEMEQKQLQ 30 Neuropeptide 1 Opioid P14 1 (Probable) 215 P12272 1 AVSEHQLLHDKGKSIQDLRRRFFLHHLIAEIHTAEIRAT 141 Parathyroid Parathyroid hormone T39 1 SEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKVET hormone-related YKEQPLKTPGKKKKGKPGKRKEQEKKKRRTRSAWLD protein SGVTGSGLEGDHLSDTSTTSLELDSRRH 217 P12272 1 ATSEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKV 57 PTHrP[38-94] Parathyroid hormone T2 1 ETYKEQPLKTPGKKKKGKP 219 P01189 1 YGGFMTSEKSQTPLVTLFKNAIIKNAYKKGE 31 Beta-endorphin POMC S10, T12, T16 3 222 P01189 1 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAAE 89 Lipotropin beta POMC S68, T70, T74 3 KKDEGPYRMEHFRWGSPPKDKRYGGFMTSEKSQTPL VTLFKNAIIKNAYKKGE 227 P01189 1 WCLESSQCQDLTTESNLLECIRACKPDLSAETPMFPGN 76 NPP POMC T45 1 GDEQPLTENPRKYVMGHFRWDRFGRRNSSSSGSSGAG Q 229 Q9UHG2 1 LETPAPQVPARRLLPP 16 Big LEN (By ProSAAS T3, P4, A5 2 similarity) 230 Q9UHG2 1 AADHDVGSELPPEGVLGALLRVKRLETPAPQVPARRL 40 Big PEN-LEN ProSAAS T27, P28, A29 2 LPP (By similarity) 231 Q9UHG2 1 ARPVKEPRGLSAASPPLAETGAPRRF 26 Big SAAS (By ProSAAS G9, S14, T20, G21, A22 5 similarity) 233 Q9UHG2 1 LETPAPQVPA 10 Little LEN (By ProSAAS T3, P4, A5 2 similarity) 234 Q9UHG2 1 GLSAASPPLAETGAPRRF 18 Little SAAS (By ProSAAS G1, S6, T12, G13, A14 5 similarity) 236 Q9UHG2 1 ARPVKEPRGLSAASPPLAETGAPRRFRRSVPRGEAAGA 227 ProSAAS ProSAAS 13 VQELARALAHLLEAERQERARAEAQEAEDQQARVLA QLLRVWGAPRNSDPALGLDDDPDAPAAQLARALLRAR G9, S14, T20, G21, A22, A82, LDPAALAAQLVPAPVPAAALRPRPPVYDDGPAGPDAE N85, S86, A143, S173, A174, EAGDETPDVDPELLRYLLGRILAGSADSEGVAAPRRLR T214, P215, A216 RAADHDVGSELPPEGVLGALLRVKRLETPAPQVPARR LLPP 252 P01019 1 DRVYIHPFHLVIHNESTCEQLAKANAGKPKDPTFIPAPI 452 Angiotensinogen Serpin T407 or S409 or T410 or T422 1 QAKTSPVDEKALQDQLVLVAAKLDTEDKLRAAMVG (Ambiguous) MLANFLGFRIYGMHSELWGVVHGATVLSPTAVFGTLA SLYLGALDHTADRLQAILGVPWKDKNCTSRLDAHKVL SALQAVQGLLVAQGRADSQAQLLLSTVVGVFTAPGLH LKQPFVQGLALYTPVVLPRSLDFTELDVAAEKIDRFMQ AVTGWKTGCSLMGASVDSTLAFNTYVHFQGKMKGFS LLAEPQEFWVDNSTSVSVPMLSGMGTFQHWSDIQDNF SVTQVPFTESACLLLIQPHYASDLDKVEGLTFQQNSLN WMKKLSPRTIHLTMPQLVLQGSYDLQDLLAQAELPAI LHTELNLQKLSNDRIRVGEVLNSIFFELEADEREPTEST QQLNKPEVLEVTLNRPFLFAVYDQSATALHFLGRVAN PLSTA 260 P61278 1 SANSNPAMAPRERKAGCKNFFWKTFTSC 28 Somatostatin-28 Somastostatin S1 or S4 (Ambiguous) 1 262 P20366 1 ALNSVAYERSAMQNYE 15 C-terminal- Tachykinin A1, S4 2 flanking peptide 267 P20396 1 QPEAAQQEAVTAAEHPGLDDFLRQVERLLFLRENIQRL 218 Pro-thyrotropin- TRH I50 1 QGDQGEHSASQIFQSDWLSKRQHPGKREEEEEEGVEE releasing hormone EEEEEGGAVGPHKRQHPGRREDEASWSVDVTQHKRQ HPGRRSPWLAYAVPKRQHPGRRLADPKAQRSWEEEEE EEEREEDLMPEKRQHPGKRALGGPCGPQGAYGQAGLL LGLLDDLSRSQGAEEKRQHPGRRAAWVREPLEE 272 P01185 1 ASDRSNATQLDGPAGALLLRLVQLAGAPEPFEPAQPDA 39 Copeptin Vasopressin/oxytocin A27 1 Y 279 O15240 1 APPGRPEAQPPPLSSEHKEPVAGDAVPGPKDGSAPEVR 591 Neurosecretory VGF P1, S12, P202, S205, 5 GARNSEPQDEGELFQGVDPRALAAVLLQALDRPASPP protein VGF T477 APSGSQQGPEEEAAEALLTETVRSQTHSLPAPESPEPAA PPRPQTPENGPEASDPSEELEALASLLQELRDFSPSSAK RQQETAAAETETRTHTLTRVNLESPGPERVWRASWGE FQARVPERAPLPPPAPSQFQARMPDSGPLPETHKFGEG VSSPKTHLGEALAPLSKAYQGVAAPFPKARRPESALLG GSEAGERLLQQGLAQVEAGRRQAEATRQAAAQEERL ADLASDLLLQYLLQGGARQRGLGGRGLQEAAEERESA REEEEAEQERRGGEERVGEEDEEAAEAEAEAEEAERA RQNALLFAEEEDGEAGAEDKRSQEETPGHRRKEAEGT EEGGEEEDDEEMDPQTIDSLIELSTKLHLPADDVVSIIEE VEEKRKRKKNAPPEPVPPPRAAPAPTHVRSPQPPPPAP APARDELPDWNEVLPPWDREEDEVYPPGPYHPFPNYIR PRTLQPPSALRRRHYHHALPPSRHYPGREAQARRAQEE AEAEERRLQEQEELENYIEHVLLRRP
TABLE-US-00011 TABLE 6E *Site position (Bold: identified or conserved SEQ Pep- Pep- Peptide in human, Italic; No. ID Entry_ tide tide Peptide Hormone identified in non-human of NO: Human Start Peptide_Sequence end Name Family and not conserved) sites 1 P05408 1 SVPHFSDEDKDPE 13 C-terminal 7B2 S1 1 peptide (By similarity) 2 P05408 1 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPA 186 Neuroendocrine 7B2 S2, T5, S10, T108, S174 5 HQAMNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAEL protein 7B2 TGDNIPKDFSEDQGYPDPPNPCPVGKTADDGCLENT PDTAEFSREFQLHQHLFDPEHDYPGLGKWNKKLLY EKMKGGERRKRRSVNPYLQGQRLDNVVAKKSVPH FSDEDKDPE 3 P05408 1 YSPRTPDRVSEADIQRLLHGVMEQLGIARPRVEYPA 150 N-terminal 7B2 S2, T5, S10, T108 4 HQAMNLVGPQSIEGGAHEGLQHLGPFGNIPNIVAEL peptide (By TGDNIPKDFSEDQGYPDPPNPCPVGKTADDGCLENT similarity) PDTAEFSREFQLHQHLFDPEHDYPGLGKWNKKLLY EKMKGGE 4 Q8N6N7 1 MALQADFDRAAEDVRKLKARPDDGELKELYGLYK 88 Acyl-CoA- ACBP QAIVGDINIACPGMLDLKGKAKWEAWNLKKGLSTE binding domain- DATSAYISKAKELIEKYGI containing protein 7 5 P07108 1 SQAEFEKAAEEVRHLKTKPSDEEMLFIYGHYKQAT 86 Acyl-CoA- ACBP T35 or T41 (ambiguous) 1 VGDINTERPGMLDFTGKAKWDAWNELKGTSKEDA binding protein MKAYINKVEELKKKYGI 6 P35318 1 YRQSMNNFQGLRSFGCRFGTCTVQKLAHQIYQFTD 52 Adrenomedullin Adrenomedullin T22, Y31, T34 3 KDKDNVAPRSKISPQGY 7 Q7Z4H4 1 TQAQLLRVGCVLGTCQVQNLSHRLWQLMGPAGRQ 47 Adrenomedullin- Adrenomedullin DSAPVDPSSPHSY 2 (By similarity) 8 Q7Z4H4 1 VGCVLGTCQVQNLSHRLWQLMGPAGRQDSAPVDP 40 Intermedin-short Adrenomedullin SSPHSY (Potential) 9 P35318 1 ARLDVASEFRKKWNKWALSR 20 Proadreno- Adrenomedullin medullin N-20 terminal peptide 10 Q9ULZ1 1 QRPRLSHKGPMPF 13 Apelin-13 (By Apelin similarity) 11 Q9ULZ1 1 NGPGPWQGGRRKFRRQRPRLSHKGPMPF 28 Apelin-28 (By Apelin similarity) 12 Q9ULZ1 1 GSRNGPGPWQGGRRKFRRQRPRLSHKGPMPF 31 Apelin-31 (By Apelin similarity) 13 Q9ULZ1 1 LVQPRGSRNGPGPWQGGRRKFRRQRPRLSHKGPMP 36 Apelin-36 (By Apelin F similarity) 14 P07492 1 VPLPAGGGTVLTKMYPRGNHWAVGHLM 27 Gastrin-releasing Bombesin/ P4, T9 2 peptide neuromedin- B/ranatensin 15 P08949 1 GNLWATGHFM 10 Neuromedin-B Bombesin/ T6 1 neuromedin- B/ranatensin 16 P08949 1 APLSWDLPEPRSRASKIRVHSRGNLWATGHFM 32 Neuromedin-B- Bombesin/ T28 1 32 neuromedin- B/ranatensin 17 P07492 1 GNHWAVGHLM 10 Neuromedin-C Bombesin/ neuromedin- B/ranatensin 18 P01042 1 RPPGFSPFR 9 Bradykinin Bradykinin 19 P01042 1 KRPPGFSPFR 10 Lysyl-bradykinin Bradykinin 20 P01042 1 ISLMKRPPGFSPFR 14 T-kinin Bradykinin 21 P01258 1 CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP 32 Calcitonin Calcitonin T6, T11, T13, T21, T25 5 22 P06881 1 ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSK 37 Calcitonin gene- Calcitonin T9, S17 2 AF related peptide 1 23 P10092 1 ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSK 37 Calcitonin gene- Calcitonin T9, S17, S25, T30, S34 5 AF related peptide 2 24 P10997 1 KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNT 37 Islet amyloid Calcitonin T5, T9, T30 3 Y polypeptide 25 P01258 1 DMSSDLERDHRPHVSMPQNAN 21 Katacalcin Calcitonin S15 1 26 Q16568 1 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKK 39 CART(1-39) CART LKS 27 Q16568 1 VPIYEKKYGQVPMCDAGEQCAVRKGARIGKLCDCP 48 CART(42-89) CART RGTSCNSFLLKCL 28 Q16568 1 QEDAELQPRALDIYSAVDDASHEKELIEALQEVLKK 89 Cocaine- and CART LKSKRVPIYEKKYGQVPMCDAGEQCAVRKGARIGK amphetamine- LCDCPRGTSCNSFLLKCL regulated 29 P23435 1 GSAKVAFSAIRSTNH 15 [des-Ser1]- Cerebellins cerebellin 30 P23435 1 SGSAKVAFSAIRSTNH 16 Cerebellin Cerebellins 31 P23435 1 QNETEPIVLEGKCLVVCDSNPTSDPTGTALGISVRSG 172 Cerebellin-1 Cerebellins SAKVAFSAIRSTNHEPSEMSNRTMIIYFDQVLVNIGN NFDSERSTFIAPRKGIYSFNFHVVKVYNRQTIQVSLM LNGWPVISAFAGDQDVTREAASNGVLIQMEKGDRA YLKLERGNLMGGWKYSTFSGFLVFPL 32 Q8IUK8 1 QNDTEPIVLEGKCLVVCDSSPSADGAVTSSLGISVRS 173 Cerebellin-2 Cerebellins GSAKVAFSATRSTNHEPSEMSNRTMTIYFDQVLVNI GNHFDLASSIFVAPRKGIYSFSFHVVKVYNRQTIQVS LMQNGYPVISAFAGDQDVTREAASNGVLLLMERED KVHLKLERGNLMGGWKYSTFSGFLVFPL 33 Q6UW01 1 QEGSEPVLLEGECLVVCEPGRAAAGGPGGAALGEA 173 Cerebellin-3 Cerebellins PPGRVAFAAVRSHHHEPAGETGNGTSGAIYFDQVL VNEGGGFDRASGSFVAPVRGVYSFRFHVVKVYNRQ TVQVSLMLNTWPVISAFANDPDVTREAATSSVLLPL DPGDRVSLRLRRGNLLGGWKYSSFSGFLIFPL 34 Q9NTU7 1 QNDTEPIVLEGKCLVVCDSNPATDSKGSSSSPLGISV 174 Cerebellin-4 Cerebellins RAANSKVAFSAVRSTNHEPSEMSNKTRIIYFDQILVN VGNFFTLESVFVAPRKGIYSFSFHVIKVYQSQTIQVN LMLNGKPVISAFAGDKDVTREAATNGVLLYLDKED KVYLKLEKGNLVGGWQYSTFSGFLVFPL 35 P10645 1 AYGFRGPGPQL 11 AL-11 Chromogranin/ secretogranin 36 P05060 1 SAEFPDFYDSEEPVSTHQEAENEKDRADQTVLTEDE 57 CCB peptide Chromogranin/ S15 1 KKELENLAAMDLELQKIAEKF secretogranin 37 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQ 439 Chromogranin-A Chromogranin/ S27, S30, S36, S80, E104, 39 ECFETLRGDERILSILRHQNLLKELQDLALQGAKER secretogranin S108, A127, T128, A135, L136, AHQQKKHSGFEDELSEVLENQSSQAELKEAVEEPSS S143, A151, N164, T165, P167, KDVMEKREDSKEAEKSGEATDGARPQALPEPMQES A169, S170, S173, E182, L188, KAEGNNQAPGEEEEEEEEATNTHPPASLPSQKYPGP S189, V193, S200, T233, L236, QAEGDSEGLSQGLVDREKGLSAEPGWQAKREEEEE N237, P238, S241, Y244, P265, EEEEAEAGEEAVPEEEGPTVVLNPHPSLGYKEIRKG E268, P273, S282, M291, V294, ESRSEALAVDGAGKPGAEEAQDPEGKGEQEHSQQK T335, A360, S380, G395 EEEEEMAVVPQGLFRGGKSGELEQEEERLSKEWEDS KRWSKMDQLAKELTAEKRLEGQEEEEDNRDSSMK LSFRARAYGFRGPGPQLRRGWRPSSREDSLEAGLPL QVRGYPEEKKEEEGSANRRPEDQELESLSAIEAELE KVAHQLQALRRG 38 P10645 1 EDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQ 92 EA-92 Chromogranin/ A12, T13, A20, L21, S28, A36, 17 APGEEEEEEEEATNTHPPASLPSQKYPGPQAEGDSEG secretogranin N49, T50, P52, A54, S55, S58, LSQGLVDREKGLSAEPGWQA E67, L73, S74, V78, S85 39 P10645 1 EEEGSANRRPEDQELESLSAIEAELEKVAHQLQALR 37 ER-37 Chromogranin/ S17 or S19 (Ambiguous) 1 R secretogranin 40 P10645 1 EEEEEEEEEAEAGEEAVPEEEGPTVVLNPHPSL 33 ES-43 Chromogranin/ T24, L27, N28, P29, S32 5 secretogranin 41 P05060 1 FLGEGHHRVQENQMDKARRHPQGAWKELDRNYLN 74 GAWK peptide Chromogranin/ H7, N12, Y35, T72, A73 5 YGEEGAPGKWQQQGDLQDTKENREEARFQDKQYS secretogranin SHHTAE 42 P10645 1 GYPEEKKEEEGSANRRPEDQELESLSAIEAELEKVA 44 GR-44 Chromogranin/ G1 1 HQLQALRR secretogranin 43 P10645 1 GWRPSSREDSLEAGLPLQV 19 GV-19 Chromogranin/ S6 1 secretogranin 44 P10645 1 LEGQEEEEDNRDSSMKLSF 19 LF-19 Chromogranin/ secretogranin 45 P10645 1 SEALAVDGAGKPGAEEAQDPEGKGEQEHSQQKEEE 48 Pancreastatin Chromogranin/ P12, E15, P20, S29, M38, V41 6 EEMAVVPQGLFRG secretogranin 46 P05060 1 MPVDNRNHNEGMVTRCIIEVLSNALSKSSAPPITPEC 657 Secretogranin-1 Chromogranin/ T14, S26, S28, S29, P32, T59, 40 RQVLKTSRKDVKDKETFENENTKFEVRLLRDPADA secretogranin S73, A75, S80, A84, E89, T96, SEAHESSSRGEAGAPGEEDIQGPTKADTEKWAEGGG S120, S124, S129, T174, N176, HSRERADEPQWSLYPSDSQVSEEVKTRHSEKSQRED S186, S273, S274, S278, P287, EEEEEGENYQKGERGEDSSEEKHLEEPGETQNAFLN S297, M298, S300, D306, T310, ERKQASAIKKEELVARSETHAAGHSQEKTHSREKSS Y328, P329, G330, K376, QESGEETGSQENHPQESKGQPRSQEESEEGEEDATS M377, H426, Y454, T491, EVDKRRTRPRHHHGRSRPDRSSQGGSLPSEEKGHPQ A492, T536, S557, N568, S611 EESEESNVSMASLGEKRDHHSTHYRASEEEPEYGEE IKGYPGVQAPEDLEWERYRGRGSEEYRAPRPQSEES WDEEDKRNYPSLELDKMAHGYGEESEEERGLEPGK GRHHRGRGGEPRAYFMSDTREEKRFLGEGHHRVQE NQMDKARRHPQGAWKELDRNYLNYGEEGAPGKW QQQGDLQDTKENREEARFQDKQYSSHHTAEKRKRL GELFNPYYDPLQWKSSHFERRDNMNDNFLEGEEEN ELTLNEKNFFPEYNYDWWEKKPFSEDVNWGYEKR NLARVPKLDLKRQYDRVAQLDQLLHYRKKSAEFPD FYDSEEPVSTHQEAENEKDRADQTVLTEDEKKELE NLAAMDLELQKIAEKFSQRG 47 P13521 1 QRNQLLQKEPDLRLENVQKFPSPEMIRALEYIENLR 587 Secretogranin-2 Chromogranin/ S22, S45, A106, Y160, T161, 16 QQAHKEESSPDYNPYQGVSVPLQQKENGDESHLPE secretogranin S164, T167, S229, T231, S348, RDSLSEEDWMRIILEALRQAENEPQSAPKENKPYAL T420, N459, G499, S526, S536, NSEKNFPMDMSDDYETQQWPERKLKHMQFPPMYE P543 ENSRDNPFKRTNEIVEEQYTPQSLATLESVFQELGK LTGPNNQKRERMDEEQKLYTDDEDDIYKANNIAYE DVVGGEDWNPVEEKIESQTQEEVRDSKENIEKNEQI NDEMKRSGQLGIQEEDLRKESKDQLSDDVSKVIAY LKRLVNAAGSGRLQNGQNGERATRLFEKPLDSQSIY QLIEISRNLQIPPEDLIEMLKTGEKPNGSVEPERELDL PVDLDDISEADLDHPDLFQNRMLSKSGYPKTPGRAG TEALPDGLSVEDILNLLGMESAANQKTSYFPNPYNQ EKVLPRLPYGAGRSRSNQLPKAAWIPHVENRQMAY ENLNDKDQELGEYLARMLVKYPEIINSNQVKRVPG QGSSEDDLQEEEQIEQAIKEHLNQGSSQETDKLAPV SKRFPVGPPKNDDTPNRQYWDEDLLMKVLEYLNQ EKAEKGREHIAKRAMENM 48 Q8WXD2 1 FPKPGGSQDKSLHNRELSAERPLNEQIAEAEEDKIKK 449 Secretogranin-3 Chromogranin/ P41, T63, S103, T104, T125, 15 TYPPENKPGQSNYSFVDNLNLLKAITEKEKIEKERQ secretogranin A138, D192, P193, T197, S198, SIRSSPLDNKLNVEDVDSTKNRKLIDDYDSTKSGLD T200, T212, T217, A222, S340 HKFQDDPDGLHQLDGTPLTAEDIVHKIAARIYEEND RAVFDKIVSKLLNLGLITESQAHTLEDEVAEVLQKLI SKEANNYEEDPNKPTSWTENQAGKIPEKVTPMAAI QDGLAKGENDETVSNTLTLTNGLERRTKTYSEDNFE ELQYFPNFYALLKSIDSEKEAKEKETLITIMKTLIDFV KMMVKYGTISPEEGVSYLENLDEMIALQTKNKLEK NATDNISKLFPAPSEKSHEETDSTKEEAAKMEKEYG SLKDSTKDDNSNPGGKTDEPKGKTEAYLEAIRKNIE WLKKHDKKGNKEDYDLSKMRDFINKQADAYVEK GILDKEEAEAIKRIYSSL 49 P13521 1 TNEIVEEQYTPQSLATLESVFQELGKLTGPNNQ 33 Secretoneurin Chromogranin/ Y9, T10, S13, T16, 4 secretogranin 50 P10645 1 SGELEQEEERLSKEWEDS 18 SS-18 Chromogranin/ secretogranin 51 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQ 76 Vasostatin-1 Chromogranin/ S27, S30, S36 3 ECFETLRGDERILSILRHQNLLKELQDLALQGAKER secretogranin AHQQ 52 P10645 1 LPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQ 113 Vasostatin-2 Chromogranin/ S27, S30, S36, S80, E104, 6 ECFETLRGDERILSILRHQNLLKELQDLALQGAKER secretogranin S108 AHQQKKHSGFEDELSEVLENQSSQAELKEAVEEPSS KDVME 53 P10645 1 WSKMDQLA 8 WA-8 Chromogranin/ secretogranin 54 P10645 1 WSKMDQLAKELTAE 14 WE-14 Chromogranin/ T12 1 secretogranin 55 P01042 1 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQ 626 Kininogen-1 Cystatin T119, T133, T243, T255, E381, 15 FVLYRIFLATKTVGSDTFYSFKYEIKEGDCPVQSGKT T382, T383, S385, T389, S390, WQDCEYKDAAKAATGECTATVGKRSSTKFSVATQ T528, S586, T592, S593, T610 TCQITPAEGPVVTAQYDCLGCVHPISTQSPDLEPILR HGIQYENNNTQHSSLEMLNEVKRAQRQVVAGLNFR ITYSIVQTNCSKENFLFLTPDCKSLWNGDTGECTDN AYIDIQLRIASFSQNCDIYPGKDEVQPPTKICVGCPR DIPTNSPELEETLTHTITKLNAENNATFYFKIDNVKK ARVQVVAGKKYFIDFVARETTCSKESNEELTESCET KKLGQSLDCNAEVYVVPWEKKIYPTVNCQPLGMIS LMKRPPGESPERSSRIGEIKEETTVSPPHTSMAPAQD EERDSGKEQGHTRRHDWGHEKQRKHNLGHGHKHE RDQGHGHQRGHGLGHGHEQQHGLGHGHKFKLDD DLEHQGGHVLDHGHKHKHGHGHGKHKNKGKKNG KHNGWKFEHLASSSEDSTTPSAQTQEKFEGPTPIPSL AKPGVTVTFSDFQDSDLIATMMPPISPAPIQSDDDWI PDIQIDPNGLSENPISDEPDTTSPKCPGRPWKSVSEIN PTTQMKESYYFDLTDGLS 56 P01042 1 QESQSEEIDCNDKDLFKAVDAALKKYNSQNQSNNQ 362 Kininogen-1 Cystatin T119, T133, T243, T255 4 FVLYRIFLATKTVGSDTFYSFKYEIKEGDCPVQSGKT heavy chain WQDCEYKDAAKAATGECTATVGKRSSTKFSVATQ TCQITPAEGPVVTAQYDCLGCVHPISTQSPDLEPILR HGIQYENNNTQHSSLEMLNEVKRAQRQVVAGLNFR ITYSIVQTNCSKENFLFLTPDCKSLWNGDTGECTDN AYIDIQLRIASFSQNCDIYPGKDEVQPPTKICVGCPR DIPTNSPELEETLTHTITKLNAENNATFYFKIDNVKK ARVQVVAGKKYFIDFVARETTCSKESNEELTESCET KKLGQSLDCNAEVYVVPWEKKIYPTVNCQPLGMIS LMK 57 P01042 1 SSRIGEIKEETTVSPPHTSMAPAQDEERDSGKEQGH 255 Kininogen-1 Cystatin E10, T11, T12, S14, T18, S19, 11 TRRHDWGHEKQRKHNLGHGHKHERDQGHGHQRG light chain T157, S215, T221, S222, T239 HGLGHGHEQQHGLGHGHKFKLDDDLEHQGGHVLD HGHKHKHGHGHGKHKNKGKKNGKHNGWKFEHL ASSSEDSTTPSAQTQEKFEGPTPIPSLAKPGVTVITS DFQDSDLIATMMPPISPAPIQSDDDWIPDIQIDPNGLS FNPISDEPDTTSPKCPGRPWKSVSEINPTTQMKESYY FDLTDGLS 58 P01042 1 WGHE 4 Low molecular Cystatin weight growth- promoting factor 59 P05305 1 CSCSSLMDKECVYFCHLDIIWVNTPEHVVPYGLGSP 38 Big endothelin-1 Endothelin/ RS sarafotoxin 60 P05305 1 CSCSSLMDKECVYFCHLDIIW 21 Endothelin-1 Endothelin/ sarafotoxin 61 P20800 1 CSCSSWLDKECVYFCHLDIIVV 21 Endothelin-2 Endothelin/ sarafotoxin 62 P14138 1 CTCFTYKDKECVYYCHLDIIW 21 Endothelin-3 Endothelin/ sarafotoxin 63 O15130 1 AGEGLNSQFVVSLAAPQRF 18 Neuropeptide AF FMRFamide related peptide 64 O15130 1 FLFQPQRF 8 Neuropeptide FF FMRFamide related peptide 65 Q9HCQ7 1 SLNFEELKDWGPKNVIKMSTPAVNKMPHSFANLPL 37 Neuropeptide FMRFamide related RF NPSF (Potential) peptide 66 Q9HCQ7 1 VPNLPQRF 8 Neuropeptide FMRFamide related NPVF peptide 67 Q9HCQ7 1 MPHSFANLPLRF 12 Neuropeptide FMRFamide related RFRP-1 peptide 68 Q9HCQ7 1 SAGATANLPLRS 12 Neuropeptide FMRFamide related RFRP-2 peptide (Potential) 69 O15130 1 SQAFLFQPQRF 11 Neuropeptide SF FMRFamide related peptide 70 P81277 1 TPDINPAWYASRGIRPVGRF 20 Prolactin- FMRFamide related releasing peptide peptide PrRP20 71 P81277 1 SRTHRHSMEIRTPDINPAWYASRGIRPVGRF 31 Prolactin- FMRFamide related releasing peptide peptide PrRP31 72 P22466 1 GWTLNSAGYLLGPHAVGNHRSFSDKNGLTS 30 Galanin Galanin T3, S23 2 73 P22466 1 ELRPEDDMKPGSFDRSIPENNIMRTIIEFLSFLHLKEA 59 Galanin Galanin A49 1 GALDRLLDLPAAASSEDIERS message- associated peptide 74 Q9UBC7 1 APAHRGRGGWTLNSAGYLLGPVLHLPQMGDQDGK 60 Galanin-like Galanin RETALEILDLWKAIDGLPYSHPPQPS peptide 75 P01350 1 QLGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF 34 Big gastrin Gastrin/ cholecystokinin 76 P06307 1 QPVPPADPAGSGLQRAEEAPRRQLRVSQRTDGESRA 95 Cholecystokinin Gastrin/ A8, T30, N65 3 HLGALLARYIQQARKAPSGRMSIVKNLQNLDPSHRI cholecystokinin SDRDYMGWMDFGRRSAEEYEYPS 77 P06307 1 ISDRDYMGWMDF 12 Cholecystokinin- Gastrin/ 12 cholecystokinin 78 P06307 1 LDPSHRISDRDYMGWMDF 18 Cholecystokinin- Gastrin/ 18 (By cholecystokinin similarity) 79 P06307 1 IVKNLQNLDPSHRISDRDYMGWMDF 25 Cholecystokinin- Gastrin/ N7 1 25 (By cholecystokinin similarity) 80 P06307 1 KAPSGRMSIVKNLQNLDPSHRISDRDYMGWMDF 33 Cholecystokinin- Gastrin/ N15 1 33 cholecystokinin 81 P06307 1 YIQQARKAPSGRMSIVKNLQNLDPSHRISDRDYMG 39 Cholecystokinin- Gastrin/ N21 1 WMDF 39 cholecystokinin 82 P06307 1 GWMDF 5 Cholecystokinin- Gastrin/ 5 (By similarity) cholecystokinin 83 P06307 1 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIV 58 Cholecystokinin- Gastrin/ T5, N40 2 KNLQNLDPSHRISDRDYMGWMDF 58 cholecystokinin 84 P06307 1 VSQRTDGESRAHLGALLARYIQQARKAPSGRMSIV 49 Cholecystokinin- Gastrin/ T5, N40 2 KNLQNLDPSHRISD 58 cholecystokinin desnonopeptide (By similarity) 85 P06307 1 YMGWMDF 7 Cholecystokinin- Gastrin/ 7 (By similarity) cholecystokinin 86 P06307 1 DYMGWMDF 8 Cholecystokinin- Gastrin/ 8 cholecystokinin 87 P01350 1 QGPWLEEEEEAYGWMDF 17 Gastrin Gastrin/ cholecystokinin 88 P01350 1 WLEEEEEAYGWMDF 14 Gastrin-14 Gastrin/ cholecystokinin 89 P01350 1 DLELPWLEQQGPASHHRRQLGPQGPPHLVADPSKK 52 Gastrin-52 Gastrin/ QGPWLEEEEEAYGWMDF cholecystokinin 90 P01350 1 YGWMDF 6 Gastrin-6 Gastrin/ cholecystokinin 91 P01350 1 SWKPRSQQPDAPLGTGANRDLELPWLEQQGPASHH 71 Gastrin-71 Gastrin/ L12 or G13 (Ambiguous) 1 RRQLGPQGPPHLVADPSKKQGPWLEEEEEAYGWM cholecystokinin DF 92 P09681 1 YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKND 42 Gastric inhibitory Glucagon WKHNITQ polypeptide 93 P01275 1 RSLQDTEEKSRSFSASQADPLSDPDQMNEDKRHSQG 69 Glicentin (By Glucagon S12, T39 2 TFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA similarity) 94 P01275 1 RSLQDTEEKSRSFSASQADPLSDPDQMNED 30 Glicentin-related Glucagon S12 1 polypeptide (By similarity) 95 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNT 29 Glucagon Glucagon T7 1 96 P01275 1 HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKG 37 Glucagon-like Glucagon T11 or T13 or S14 1 RG peptide 1 (Ambiguous) 97 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR 31 Glucagon-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1(7-36) 98 P01275 1 HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG 30 Glucagon-like Glucagon T5 or T7 or S8 (Ambiguous) 1 peptide 1(7-37) 99 P01275 1 HADGSFSDEMNTILDNLAARDFINWLIQTKITD 33 Glucagon-like Glucagon peptide 2 (By similarity) 100 P01282 1 HADGVFTSDFSKLLGQLSAKKYLESLM 27 Intestinal peptide Glucagon T7, S8 2 PHM-27 101 P01282 1 HADGVFTSDFSKLLGQLSAKKYLESLMGKRVSSNIS 42 Intestinal peptide Glucagon T7, S8 2 EDPVPV PHV-42 102 P01275 1 HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRN 37 Oxyntomodulin Glucagon T7 1 NIA (By similarity) 103 P18509 1 DVAHGILNEAYRKVLDQLSAGKHLQSLVARGVGGS 48 PACAP-related Glucagon LGGGAGDDAEPLS peptide 104 P18509 1 HSDGIFTDSYSRYRKQMAVKKYLAAVL 27 Pituitary Glucagon adenylate cyclase- activating polypeptide 27 105 P18509 1 HSDGIFTDSYSRYRKQMAVKKYLAAVLGKRYKQR 38 Pituitary Glucagon VKNK adenylate cyclase- activating polypeptide 38 106 P09683 1 HSDGTFTSELSRLREGARLQRLLQGLV 27 Secretin Glucagon T7, S8, S11 3 107 P01286 1 YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESN 44 Somatoliberin Glucagon QERGARARL 108 P01282 1 HSDAVFTDNYTRLRKQMAVKKYLNSILN 28 Vasoactive Glucagon T7 1 intestinal peptide 109 P01148 1 DAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLR 56 GnRH-associated GnRH T28 1 DLKGALESLIEEETGQKKI peptide 1 110 O43555 1 ALSSAQDPQNALRPPGRALDTAAGSPVQTAHGLPS 84 GnRH-associated GnRH DALAPLDDSMPWEGRTTAQWSLHRKRHLARTLLT peptide 2 AAREPRPAPPSSNKV 111 P01148 1 QHWSYGLRPG 10 Gonadoliberin-1 GnRH 112 O43555 1 QHWSHGWYPG 10 Gonadoliberin-2 GnRH 113 P01148 1 QHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAET 69 Progonadoliberin- GnRH T41 1 QRFECTTHQPRSPLRDLKGALESLIEEETGQKKI 1 114 O43555 1 QHWSHGWYPGGKRALSSAQDPQNALRPPGRALDT 97 Progonadoliberin- GnRH AAGSPVQTAHGLPSDALAPLDDSMPWEGRTTAQW 2 SLHRKRHLARTLLTAAREPRPAPPSSNKV 115 P01308 1 GIVEQCCTSICSLYQLENYCN 21 Insulin A chain Insulin 116 P01308 1 FVNQHLCGSHLVEALYLVCGERGFFYTPKT 30 Insulin B chain Insulin T27 (Ambiguous) 1 117 P05019 1 GPETLCGAELVDALQFVCGDRGFYFNKPTGYGSSSR 70 Insulin-like Insulin RAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA growth factor I 118 P01344 1 AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVS 67 Insulin-like Insulin S50, T62 2 RRSRGIVEECCFRSCDLALLETYCATPAKSE growth factor II 119 P01344 1 YRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSR 66 Insulin-like Insulin S49, T61 2 RSRGIVEECCFRSCDLALLETYCATPAKSE growth factor II Ala-25 Del 120 P01344 1 DVSTPPTVLPDNFPRYPVGKFFQYDTWKQSTQRL 34 Preptin Insulin S3, T4, T7 3 121 P04090 1 QLYSALANKCCHVGCTKRSLARFC 24 Relaxin A chain Insulin 122 P04808 1 PYVALFEKCCLIGCTKRSLAKYC 23 Relaxin A chain Insulin (By similarity) 123 P04090 1 DSWMEEVIKLCGRELVRAQIAICGMSTWS 29 Relaxin B chain Insulin 124 P04808 1 VAAKWKDDVIKLCGRELVRAQIAICGMSTWS 31 Relaxin B chain Insulin (By similarity) 125 Q8WXF3 1 DVLAGLSSSCCKWGCSKSEISSLC 24 Relaxin-3 A Insulin chain (By similarity) 126 Q8WXF3 1 RAAPYGVRLCGREFIRAVIFTCGGSRW 27 Relaxin-3 B Insulin chain (By similarity) 127 Q15726 1 YNWNSFGLRF 10 Kisspeptin-10 KISS1 128 Q15726 1 LPNYNWNSFGLRF 13 Kisspeptin-13 KISS1 129 Q15726 1 DLPNYNWNSFGLRF 14 Kisspeptin-14 KISS1 130 Q15726 1 EPLEKVASVGNSRPTGQQLESLGLLAPGEQSLPCIL 119 Metastasis- KISS1 A69 1 RKPAATARLSRRGTSLSPPPESSGSPQQPGLSAPHSR suppressor KiSS- QIPAPQGAVLVQREKDLPNYNVVNSFGLRFGKREAA 1 PGNHGRSAGRG 131 Q15726 1 GTSLSPPPESSGSPQQPGLSAPHSRQIPAPQGAVLVQ 54 Metastin KISS1 A21 1 REKDLPNYNWNSFGLRF 132 P41159 1 VPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVT 146 Leptin Leptin GLDFIPGLHPILTLSKMDQTLAVYQQILTSMPSRNVI QISNDLENLRDLLHVLAFSKSCHLPWASGLETLDSL GGVLEASGYSTEVVALSRLQGSLQDMLWQLDLSPG C 133 P20382 1 DFDMLRCMLGRVYRPCWQV 19 Melanin- Melanin- concentrating concentrating hormone hormone 134 P20382 1 EIGDEENSAKFPI 13 Neuropeptide- Melanin- glutamic acid- concentrating isoleucine hormone 135 P20382 1 GSVAFPAENGVQNTESTQE 19 Neuropeptide- Melanin- T14, T16, T17 3 glycine-glutamic concentrating acid (Potential) hormone 136 P20382 1 ILLSASKSIRNLDDDMVFNTFRLGKGFQKEDTAEKS 144 Pro-MCH Melanin- S36, S41, N72, T102, T104, 6 VIAPSLEQYKNDESSFMNEEENKVSKNTGSKHNFLN concentrating T105 HGLPLNLAIKPYLALKGSVAFPAENGVQNTESTQEK hormone REIGDEENSAKFPIGRRDFDMLRCMLGRVYRPCWQ V 137 Q9UBU3 1 GSSFLSPEHQRVQQRKESKKPPAKLQP 27 Ghrelins-27 Motilin 138 Q9UBU3 1 GSSFLSPEHQRVQQRKESKKPPAKLQPR 28 Ghrelins-28 Motilin 139 P12872 1 FVPIFTYGELQRMQEKERNKGQ 22 Motilin Motilin 140 P12872 1 SLSVWQRSGEEGPVDPAEPIREEENEMIKLTAPLEIG 66 Motilin- Motilin MRMNSRQLEKYPATLEGLLSEMLPQHAAK associated peptide 141 Q9UBU3 1 FNAPFDVGIKLSGVQYQQHSQAL 23 Obestatin Motilin 142 P12872 1 FVPIFTYGELQRMQEKERNKGQKKSLSVWQRSGEE 90 Promotilin Motilin GPVDPAEPIREEENEMIKLTAPLEIGMRMNSRQLEK YPATLEGLLSEMLPQHAAK 143 Q15848 1 ETTTQGPGVLLPLPKGACTGWMAGIPGHPGHNGAP 226 Adiponectin NA T4 1 GRDGRDGTPGEKGEKGDPGLIGPKGDIGETGVPGAE GPRGFPGIQGRKGEPGEGAYVYRSAFSVGLETYVTI PNMPIRETKIFYNQQNHYDGSTGKEHCNIPGLYYFA YHITVYMKDVKVSLFKKDKAMLFTYDQYQENNVD QASGSVLLHLEVGDQVWLQVYGEGERNGLYADND NDSTFTGFLLYHDTN 144 O00253 1 AQMGLAPMEGIRRPDQALLPELPGLGLRAPLKKTT 111 Agouti-related NA T35 1 AEQAEEDLLQEAQALAEVLDLQDREPRSSRRCVRL protein HESCLGQQVPCCDPCATCYCRFFNAFCYCRKLGTA MNPCSRT 145 Q9BZL1 1 MIEVVCNDRLGKKVRVKCNTDDTIGDLKKLIAAQT 73 Ubiquitin-like NA GTRWNKIVLKKWYTIFKDHVSLGDYEIHDGMNLEL protein 5 YYQ 146 P43490 1 MNPAAEAEFNILLATDSYKVTHYKQYPPNTSKVYS 491 Nicotinamide NAPRTase YFECREKKTENSKLRKVKYEETVEYGLQYILNKYL phosphoribosyltr KGKVVTKEKIQEAKDVYKEHFQDDVFNEKGWNYI ansferase LEKYDGHLPIEIKAVPEGFVIPRGNVLFTVENTDPEC YWLTNWIETILVQSWYPITVATNSREQKKILAKYLL ETSGNLDGLEYKLHDFGYRGVSSQETAGIGASAHL VNFKGTDTVAGLALIKKYYGTKDPVPGYSVPAAEH STITAWGKDHEKDAFEHIVTQFSSVPVSVVSDSYDI YNACEKIWGEDLRHLIVSRSTQAPLIIRPDSGNPLDT VLKVLEILGKKFPVTENSKGYKLLPPYLRVIQGDGV DINTLQEIVEGMKQKMVVSIENIAFGSGGGLLQKLTR DLLNCSEKCSYVVTNGLGINVEKDPVADPNKRSKK GRLSLHRTPAGNFVTLEEGKGDLEEYGQDLLHTVF KNGKVTKSYSFDEIRKNAQLNIELEAAHH 147 P01160 1 SLRRSSCFGGRMDRIGAQSGLGCNSFRY 28 Atrial natriuretic Natriuretic peptide S19, S25 2 factor 148 P16860 1 SPKMVQGSGCFGRKMDRISSSSGLGCKV 28 BNP(1-28) Natriuretic peptide 149 P16860 1 SPKMVQGSGCFGRKMDRISSSSGLGCKVL 29 BNP(1-29) Natriuretic peptide 150 P16860 1 SPKMVQGSGCFGRKMDRISSSSGLGCKVLR 30 BNP(1-30) Natriuretic peptide 151 P16860 1 PKMVQGSGCFGRKMDRISSSSGLGCKVLRR 30 BNP(2-31) Natriuretic peptide 152 P16860 1 KMVQGSGCFGRKMDRISSSSGLGCKVL 27 BNP(3-29) Natriuretic peptide 153 P16860 1 KMVQGSGCFGRKMDRISSSSGLGCKVLR 28 BNP(3-30) Natriuretic peptide 154 P16860 1 KMVQGSGCFGRKMDRISSSSGLGCKVLRRH 30 BNP(3-32) Natriuretic peptide 155 P16860 1 MVQGSGCFGRKMDRISSSSGLGCK 24 BNP(4-27) Natriuretic peptide 156 P16860 1 MVQGSGCFGRKMDRISSSSGLGCKVL 26 BNP(4-29) Natriuretic peptide 157 P16860 1 MVQGSGCFGRKMDRISSSSGLGCKVLR 27 BNP(4-30) Natriuretic peptide 158 P16860 1 MVQGSGCFGRKMDRISSSSGLGCKVLRR 28 BNP(4-31) Natriuretic peptide 159 P16860 1 MVQGSGCFGRKMDRISSSSGLGCKVLRRH 29 BNP(4-32) Natriuretic peptide 160 P16860 1 VQGSGCFGRKMDRISSSSGLGCKVL 25 BNP(5-29) Natriuretic peptide 161 P16860 1 VQGSGCFGRKMDRISSSSGLGCKVLRR 27 BNP(5-31) Natriuretic peptide 162 P16860 1 VQGSGCFGRKMDRISSSSGLGCKVLRRH 28 BNP(5-32) Natriuretic peptide 163 P16860 1 SPKIVIVQGSGCFGRKMDRISSSSGLGCKVLRRH 32 Brain natriuretic Natriuretic peptide peptide 32 164 P01160 1 NPMYNAVSNADLMDFKNLLDHLEEKMPLED 30 Cardiodilatin- Natriuretic peptide Y4, A6, S8 (Ambiguous) 1 related peptide 165 P23582 1 GLSKGCFGLKLDRIGSMSGLGC 22 CNP-22 Natriuretic peptide 166 P23582 1 YKGANKKGLSKGCFGLKLDRIGSMSGLGC 29 CNP-29 Natriuretic peptide 167 P23582 1 DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLS 53 CNP-53 Natriuretic peptide KGCFGLKLDRIGSMSGLGC 168 P16860 1 HPLGSPGSASDLETSGLQEQRNHLQGKLSELQVEQT 108 Natriuretic Natriuretic peptide T48, E59 2 SLEPLQESPRPTGVWKSREVATEGIRGHRKMVLYTL peptides B RAPRSPKMVQGSGCFGRKMDRISSSSGLGCKVLRR H 169 P58417 1 ANLTNGGKSELLKSGSSKSTLKHIWTESSKDLSISRL 250 Neurexophilin-1 Neurexophilin LSQTFRGKENDTDLDLRYDTPEPYSEQDLWDWLRN STDLQEPRPRAKRRPIVKTGKFKMFGWGDFHSNIK TVKLNLLITGKIVDHGNGTFSVYFRHNSTGQGNVSV SLVPPTKIVEFDLAQQTVIDAKDSKSFNCRIEYEKVD KATKNTLCNYDPSKTCYQEQTQSHVSWLCSKPFKV ICIYISFYSTDYKLVQKVCPDYNYHSDTPYFPSG 170 O95156 1 KEVVHATEGLDWEDKDAPGTLVGNVVHSRIISPLR 242 Neurexophilin-2 Neurexophilin T7 1 LFVKQSPVPKPGPMAYADSMENFWDWLANITEIQE PLARTKRRPIVKTGKFKKMFGWGDFHSNIKTVKLN LLITGKIVDHGNGTFSVYFRHNSTGLGNVSVSLVPPS KVVEFEVSPQSTLETKESKSFNCRIEYEKTDRAKKT ALCNFDPSKICYQEQTQSHVSWLCSKPFKVICIYIAF YSVDYKLVQKVCPDYNYHSETPYLSSG 171 O95157 1 QDDGPPGSEDPERDDHEGQPRPRVPRKRGHISPKSR 230 Neurexophilin-3 Neurexophilin PMANSTLLGLLAPPGEAWGILGQPPNRPNHSPPPSA KVKKIFGWGDFYSNIKTVALNLLVTGKIVDHGNGT FSVHFQHNATGQGNISISLVPPSKAVEFHQEQQIFIEA KASKIFNCRMEWEKVERGRRTSLCTHDPAKICSRDH AQSSATWSCSQPFKVVCVYIAFYSTDYRLVQKVCP DYNYHSDTPYYPSG 172 O95158 1 QIPESGRPQYLGLRPAAAGAGAPGQQLPEPRSSDGL 285 Neurexophilin-4 Neurexophilin GVGRAWSWAWPTNHTGALARAGAAGALPAQRTK RKPSIKAARAKKIFGWGDFYFRVHTLKFSLLVTGKI VDHVNGTFSVYFRHNSSSLGNLSVSIVPPSKRVEFG GVWLPGPVPHPLQSTLALEGVLPGLGPPLGMAAAA AGPGLGGSLGGALAGPLGGALGVPGAKESRAFNCH VEYEKTNRARKHRPCLYDPSQVCFTEHTQSQAAWL CAKPFKVICIFVSFLSFDYKLVQKVCPDYNFQSEHPY FG 173 Q5H8A3 1 ILQRGSGTAAVDFTKKDHTATWGRPFFLFRPRN 33 Neuromedin-S Neuromedins 174 P48645 1 FRVDEEFQSPFASQSRGYFLFRPRN 25 Neuromedin-U- Neuromedins 25 175 Q8NG41 1 WYKPAAGHSSYSVGRAAGLLSGLR 24 Neuropeptide B- Neuromedins 23 176 Q8NG41 1 WYKPAAGHSSYSVGRAAGLLSGLRRSPYA 29 Neuropeptide B- Neuromedins 29 177 POC0P6 1 SFRNGVGTGMKKTSFQRAKS 20 Neuropeptide S Neuromedins 178 Q8N729 1 WYKHVASPRYHTVGRAAGLLMGL 23 Neuropeptide W- Neuromedins 23 179 Q8N729 1 WYKHVASPRYHTVGRAAGLLMGLRRSPYLW 30 Neuropeptide W- Neuromedins 30 180 P30990 1 SDSEEEMKALEADFLTNMHTSKISKAHVPSWKMTL 125 Large Neurotensin LNVCSLVNNLNSPAEETGEVHEEELVARRKLPTALD neuromedin N GFSLEAMLTIYQLHKICHSRAFQHWELIQEDILDTGN DKNGKEEVIKRKIPYIL 181 P30990 1 IPYIL 5 Neuromedin N Neurotensin 182 P30990 1 QLYENKPRRPYIL 13 Neurotensin Neurotensin 183 P30990 1 DSYYY 5 Tail peptide Neurotensin (Potential) 184 P01303 1 SSPETLISDLLMRESTENVPRTRLEDPAMW 30 C-flanking NPY S1, S2, T16, T21, A28 3 peptide of NPY 185 P01303 1 YPSKPDNPGEDAPAEDMARYYSALRHYINLITRQRY 36 Neuropeptide Y NPY S3, T32 2 186 P01298 1 APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPR 36 Pancreatic NPY T32 1 Y hormone 187 P01298 1 HKEDTLAFSEWGSPHAAVPR 20 Pancreatic NPY icosapeptide 188 P10082 1 YPIKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 36 Peptide YY NPY T32 1 189 P10082 1 IKPEAPREDASPEELNRYYASLRHYLNLVTRQRY 34 Peptide YY (3- NPY T30 1 36) 190 P80303 1 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQV 82 Nesfatin-1 Nucleobindin T8, I13, S18, T26, Y29, Y30, 8 IDVLETDKHFREKLQKADIEEIKSGRLSKELDLVSHH T43, S72 VRTKLDEL 191 Q02818 1 VPLERGAPNKEETPATESPDTGLYYHRYLQEVIDVL 435 Nucleobindin-1 Nucleobindin P8, T13, T16, S18, T21, Y24, 22 ETDGHFREKLQAANAEDIKSGKLSRELDFVSHHVRT S67, T122, T136, S198, S294, KLDELKRQEVSRLRMLLKAKMDAEQDPNVQVDHL S295, T309, H313, S343, T346, NLLKQFEHLDPQNQHTFEARDLELLIQTATRDLAQY S352, Q381, A388, T400, L426, DAAHHEEFKRYEMLKEHERRRYLESLGEEQRKEAE x433(deleterious mutation) RKLEEQQRRHREHPKVNVPGSQAQLKEVWEELDGL DPNRFNPKTFFILHDINSDGVLDEQELEALFTKELEK VYDPKNEEDDMREMEEERLRMREHVMKNVDTNQ DRLVTLEEFLASTQRKEFGDTGEGWETVEMHPAYT EEELRRFEEELAAREAELNAKAQRLSQETEALGRSQ GRLEAQKRELQQAVLHMEQRKQQQQQQQGHKAP AAHPEGQLKFHPDTDDVPVPAPAGDQKEVDTSEKK LLERLPEVEVPQHL 192 P80303 1 VPIDIDKTKVQNIHPVESAKIEPPDTGLYYDEYLKQV 396 Nucleobindin-2 Nucleobindin T8, I13, S18, T26, Y29, Y30, 16 IDVLETDKHFREKLQKADIEEIKSGRLSKELDLVSHH T43, S72, S128, T139, T148, VRTKLDELKRQEVGRLRMLIKAKLDSLQDIGMDHQ A331, Y365, P382, S384, L390 ALLKQFDHLNHLNPDKEESTDLDMLIKAATSDLEH YDKTRHEEFKKYEMMKEHERREYLKTLNEEKRKE EESKILEMKKKHENHPKVNHPGSKDQLKEVWEET DGLDPNDFDPKTFFKLHDVNSDGFLDEQELEALFTK ELEKVYDPKNEEDDMVEMEEERLRMREHVMSEVD TNKDRLVTLEEFLKATEKKEFLEPDSWETLDQQQFF TEEELKEYENHALQENELKKKADELQKQKEELQRQ HDQLEAQKLEYHQVIQQMEQKKLQQGIPPSGPAGE LKFEPHI 193 P01213 1 YGGFLRKYPK 10 Alpha- Opioid neoendorphin 194 P01213 1 YGGFLRKYP 9 Beta- Opioid neoendorphin 195 P01213 1 YGGFLRRIRPKLKWDNQKRYGGFLRRQFKVVT 32 Big dynorphin Opioid 196 P01213 1 YGGFLRRIRPKLK 13 Dynorphin A(1- Opioid 13) (By similarity) 197 P01213 1 YGGFLRRIRPKLKWDNQ 17 Dynorphin A(1- Opioid 17) 198 P01213 1 YGGFLRRI 8 Dynorphin A(1- Opioid 8) (By similarity) 199 P01213 1 YGGFL 5 Leu-enkephalin Opioid 200 P01213 1 YGGFLRRQFKVVTRSQEDPNAYSGELFDA 29 Leumorphin Opioid 201 P01210 1 YGGFM 5 Met-enkephalin Opioid 202 P01210 1 YGGFMRGL 8 Met-enkephalin- Opioid Arg-Gly-Leu 203 P01210 1 YGGFMRF 7 Met-enkephalin- Opioid Arg-Phe 204 Q13519 1 MPRVRSLFQEQEEPEPGMEEAGEMEQKQLQ 30 Neuropeptide 1 Opioid P14 1 (Probable) 205 Q13519 1 FSEFMRQYLVLSMQSSQ 17 Neuropeptide 2 Opioid (Probable) 206 Q13519 1 FGGFTGARKSARKLANQ 17 Nociceptin Opioid 207 P01210 1 DAEEDDSLANSSDLLKELLETGDNRERSHHQDGSD 41 PENK(143-183) Opioid NEEEVS (By similarity) 208 P01210 1 FAEALPSDEEGESYSKEVPEME 22 PENK(237-258) Opioid (By similarity) 209 P01213 1 YGGFLRRQFKVVT 13 Rimorphin Opioid 210 P01210 1 MDELYPMEPEEEANGSEILA 20 rimorphin Opioid 211 P01210 1 ECSQDCATCSYRLVRPADINFLACVMECEGKLPSLK 73 Synenkephalin Opioid IWETCKELLQLSKPELPQDGTSTLRENSKPEESHLLA 212 O43612 1 QPLPDCCRQKTCSCRLYELLHGAGNHAAGILTL 33 Orexin-A Orexin 213 O43612 1 RSGPPGLQGRLQRLLQASGNHAAGILTM 28 Orexin-B Orexin 214 P12272 1 TRSAWLDSGVTGSGLEGDHLSDTSTTSLELDSR 33 Osteostatin Parathyroid hormone 215 P12272 1 AVSEHQLLHDKGKSIQDLRRREFLHHLIAEIHTAEIR 141 Parathyroid Parathyroid hormone T39 1 ATSEVSPNSKPSPNTKNHPVREGSDDEGRYLTQETN hormone-related KVETYKEQPLKTPGKKKKGKPGKRKEQEKKKRRTR protein SAWLDSGVTGSGLEGDHLSDTSTTSLELDSRRH 216 P12272 1 AVSEHQLLHDKGKSIQDLRRREFLHHLIAEIHTAEI 36 PTHrP[1-36] Parathyroid hormone 217 P12272 1 ATSEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETN 57 PTHrP[38-94] Parathyroid hormone T2 1 KVETYKEQPLKTPGKKKKGKP 218 Q96A98 1 SLALADDAAFRERARLLAALERRHWLNSYMHKLL 39 Tuberoinfundibular Parathyroid hormone VLDAP peptide of 39residues 219 P01189 1 YGGFMTSEKSQTPLVTLEKNAIIKNAYKKGE 31 Beta-endorphin POMC S10, T12, T16 3 220 P01189 1 SYSMEHFRWGKPVGKKRRPVKVYPNGAEDESAEA 39 Corticotropin POMC FPLEF 221 P01189 1 PVKVYPNGAEDESAEAFPLEF 21 Corticotropin- POMC like intermediary peptide 222 P01189 1 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAA 89 Lipotropin beta POMC S68, T70, T74 3 EKKDEGPYRMEHFRWGSPPKDKRYGGFMTSEKSQ TPLVTLFKNAIIKNAYKKGE 223 P01189 1 ELTGQRLREGDGPDGPADDGAGAQADLEHSLLVAA 56 Lipotropin POMC EKKDEGPYRMEHFRWGSPPKD gamma 224 P01189 1 SYSMEHFRWGKPV 13 Melanotropin POMC alpha 225 P01189 1 DEGPYRMEHFRWGSPPKD 18 Melanotropin POMC beta 226 P01189 1 YVMGHFRWDRF 11 Melanotropin POMC gamma 227 P01189 1 WCLESSQCQDLTTESNLLECIRACKPDLSAETPMFP 76 NPP POMC T45 1 GNGDEQPLTENPRKYVMGHFRWDRFGRRNSSSSGS SGAGQ 228 P01189 1 EDVSAGEDCGPLPEGGPEPRSDGAKPGPRE 30 Potential peptide POMC 229 Q9UHG2 1 LETPAPQVPARRLLPP 16 Big LEN (By ProSAAS T3, P4, A5 2 similarity) 230 Q9UHG2 1 AADHDVGSELPPEGVLGALLRVKRLETPAPQVPAR 40 Big PEN-LEN ProSAAS T27, P28, A29 2 RLLPP (By similarity) 231 Q9UHG2 1 ARPVKEPRGLSAASPPLAETGAPRRF 26 Big SAAS (By ProSAAS G9, S14, T20, G21, A22 5 similarity) 232 Q9UHG2 1 ARPVKEP 7 KEP (By ProSAAS similarity) 233 Q9UHG2 1 LETPAPQVPA 10 Little LEN (By ProSAAS T3, P4, A5 2 similarity) 234 Q9UHG2 1 GLSAASPPLAETGAPRRF 18 Little SAAS (By ProSAAS G1, S6, T12, G13, A14 5 similarity) 235 Q9UHG2 1 AADHDVGSELPPEGVLGALLRV 22 PEN (By ProSAAS similarity) 236 Q9UHG2 1 ARPVKEPRGLSAASPPLAETGAPRRFRRSVPRGEAA 227 ProSAAS ProSAAS G9, S14, T20, G21, A22, A82, 13 GAVQELARALAHLLEAERQERARAEAQEAEDQQA N85, S86, A143, S173, A174, RVLAQLLRVWGAPRNSDPALGLDDDPDAPAAQLAR T214, P215, A216 ALLRARLDPAALAAQLVPAPVPAAALRPRPPVYDD GPAGPDAEEAGDETPDVDPELLRYLLGRILAGSADS EGVAAPRRLRRAADHDVGSELPPEGVLGALLRVKR LETPAPQVPARRLLPP 237 Q9HD89 1 KTLCSMEEAINERIQEVAGSLIFRAISSIGLECQSVTS 90 Resistin Resistin/FIZZ RGDLATCPRGFAVTGCTCGSACGSWDVRAETTCHC QCAGMDWTGARCCRVQP 238 Q9BQ08 1 QCSLDSVMDKKIKDVLNSLEYSPSPISKKLSCASVKS 88 Resistin-like beta Resistin/FIZZ QGRPSSCPAGMAVTGCACGYGCGSWDVQLETTCH CQCSVVDWTTARCCHLT 239 P83859 1 QDEGSEATGFLPAAGEKTSGPLGNLAEELNGYSRK 43 QRF-amide RFamide neuropeptide KGGFSFRF 240 P06850 1 SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRK 41 Corticoliberin Sauvagine/ LMEII corticotropin- releasing factor/urotensin I 241 P55089 1 DNPSLSIDLTFHLLRTLLELARTQSQRERAEQNRIIFD 40 Urocortin Sauvagine/ SV corticotropin- releasing factor/urotensin I 242 Q96RP3 1 IVLSLDVPIGLLQILLEQARARAAREQATTNARILAR 41 Urocortin-2 Sauvagine/ VGHC corticotropin- releasing factor/urotensin I 243 Q969E3 1 FTLSLDVPTNIMNLLFNIAKAKNLRAQAAANAHLM 38 Urocortin-3 Sauvagine/ AQI corticotropin- releasing factor/urotensin I 244 P01019 1 DRVY 4 Angiotensin 1-4 Serpin 245 P01019 1 DRVYI 5 Angiotensin 1-5 Serpin 246 P01019 1 DRVYIHP 7 Angiotensin 1-7 Serpin 247 P01019 1 DRVYIHPFH 9 Angiotensin 1-9 Serpin 248 P01019 1 DRVYIHPFHL 10 Angiotensin-1 Serpin 249 P01019 1 DRVYIHPF 8 Angiotensin-2 Serpin 250 P01019 1 RVYIHPF 7 Angiotensin-3 Serpin 251 P01019 1 VYIHPF 6 Angiotensin-4 Serpin 252 P01019 1 DRVYIHPFHLVIHNESTCEQLAKANAGKPKDPTFIPA 452 Angiotensinogen Serpin T407 or S409 or T410 or T422 1 PIQAKTSPVDEKALQDQLVLVAAKLDFEDKLRAAM (Ambiguous) VGMLANFLGFRIYGMHSELWGVVHGATVLSPTAVF GTLASLYLGALDHTADRLQAILGVPWKDKNCTSRL DAHKVLSALQAVQGLLVAQGRADSQAQLLLSTVV GVFTAPGLHLKQPFVQGLALYTPVVLPRSLDFTELD VAAEKIDRFMQAVTGWKTGCSLMGASVDSTLAFN TYVHFQGKMKGFSLLAEPQEFWVDNSTSVSVPMLS GMGTFQHWSDIQDNFSVTQVPFTESACLLLIQPHYA SDLDKVEGLTFQQNSLNWMKKLSPRTIHLTMPQLV LQGSYDLQDLLAQAELPAILHFELNLQKLSNDRIRV GEVLNSIFFELEADEREPTESTQQLNKPEVLEVTLNR PFLFAVYDQSATALHFLGRVANPLSTA 253 P08185 1 MDPNAAYVNMSNHHRGLASANVDFAFSLYKHLVA 383 Corticosteroid- Serpin LSPKKNIFISPVSISMALAMLSLGTCGHTRAQLLQGL binding globulin GFNLTERSETEIHQGFQHLHQLFAKSDTSLEMTMGN ALFLDGSLELLESFSADIKHYYESEVLAMNFQDWAT ASRQINSYVKNKTQGKIVDLFSGLDSPAILVLVNYIF FKGTWTQPFDLASTREENFYVDETTVVKVPMMLQS STISYLHDSELPCQLVQMNYVGNGTVFFILPDKGKM NTVIAALSRDTINRWSAGLTSSQVDLYIPKVTISGVY DLGDVLEEMGIADLFTNQANFSRITQDAQLKSSKVV HKAVLQLNEEGVDTAGSTGVTLNLTSKPIILRFNQPF IIMIFDHFTWSSLFLARVMNPV 254 Q86U17 1 QPLLAHGDKSLQGPQPPRHQLSEPAPAYHRITPTITN 403 Serpin A11 Serpin FALRLYKELAADAPGNIFFSPVSISTTLALLSLGAQA NTSALILEGLGFNLTETPEADIHQGFRSLLHTLALPSP KLELKVGNSLFLDKRLKPRQHYLDSIKELYGAFAFS ANFTDSVTTGRQINDYLRRQTYGQVVDCLPEFSQDT FMVLANYIFFKAKWKHPFSRYQTQKQESFFVDERTS LQVPMMHQKEMHRFLYDQDLACTVLQIEYRGNAL ALLVLPDPGKMKQVEAALQPQTLRKWGQLLLPSLL DLHLPRFSISGTYNLEDILPQIGLTNILNLEADFSGVT GQLNKTISKVSHKAMVDMSEKGTEAGAASGLLSQP PSLNTMSDPHAHFNRPFLLLLWEVTTQSLLFLGKVV NPVAG 255 Q8IW75 1 LKPSFSPRNYKALSEVQGWKQRMAAKELARQNMD 394 Serpin A12 Serpin LGFKLLKKLAFYNPGRNIFLSPLSISTAFSMLCLGAQ DSTLDEIKQGFNFRKMPEKDLHEGFHYIIHELTQKT QDLKLSIGNTLFIDQRLQPQRKFLEDAKNFYSAETIL TNFQNLEMAQKQINDFISQKTHGKINNLIENIDPGTV MLLANYIFFRARWKHEFDPNVTKEEDFFLEKNSSVK VPMMFRSGIYQVGYDDKLSCTILEIPYQKNITAIFILP DEGKLKHLEKGLQVDTFSRWKTLLSRRVVDVSVPR LHMTGTFDLKKTLSYIGVSKIFEEHGDLTKIAPHRSL KVGEAVHKAELKMDERGTEGAAGTGAQTLPMETP LVVKIDKPYLLLIYSEKIPSVLFLGKIVNPIGK 256 O75830 1 SRCSAQKNTEFAVDLYQEVSLSHKDNIIFSPLGITLV 387 Serpin 12 Serpin LEMVQLGAKGKAQQQIRQTLKQQETSAGEEFFVLK SFFSAISEKKQEFTFNLANALYLQEGFTVKEQYLHG NKEFFQSAIKLVDFQDAKACAEMISTWVERKTDGKI KDMFSGEEFGPLTRLVLVNAIYFKGDWKQKFRKED TQLINFTKKNGSTVKIPMMKALLRTKYGYFSESSLN YQVLELSYKGDEFSLIIILPAEGMDIEEVEKLITAQQI LKWLSEMQEEEVEISLPREKVEQKVDEKDVLYSLNI TEIFSGGCDLSGITDSSEVYVSQVTQKVFFEINEDGS EAATSTGIHIPVIMSLAQSQFIANHPFLFIMKHNPTES ILFMGRVTNPDTQEIKGRDLDSL 257 O00230 1 DRMPCRNFFWKTFSSCK 17 Cortistatin-17 Somastostatin 258 O00230 1 QEGAPPQQSARRDRMPCRNFFWKITSSCK 29 Cortistatin-29 Somastostatin (Potential) 259 P61278 1 AGCKNFFWKTFTSC 14 Somatostatin-14 Somastostatin 260 P61278 1 SANSNPAMAPRERKAGCKNFFWKTFTSC 28 Somatostatin-28 Somastostatin S1 or S4 (Ambiguous) 1 261 P01236 1 LPICPGGAARCQVTLRDLFDRAVVLSHYIHNLSSEM 199 Prolactin Somatotropin/ FSEFDKRYTHGRGFITKAINSCHTSSLATPEDKEQAQ prolactin QMNQKDFLSLIVSILRSWNEPLYHLVTEVRGMQEAP EAILSKAVEIEEQTKRLLEGMELIVSQVHPETKENEI YPVWSGLPSLQMADEESRLSAYYNLLHCLRRDSHK IDNYLKLLKCRIIHNNNC 262 P20366 1 ALNSVAYERSAMQNYE 15 C-terminal- Tachykinin A1, S4 2 flanking peptide 263 P20366 1 HKTDSFVGLM 10 Neurokinin A Tachykinin 264 Q9UHF0 1 DMHDFFVGLM 10 Neurokinin-B Tachykinin 265 P20366 1 DADSSIEKQVALLKALYGHGQISHKRHKTDSFVGL 36 Neuropeptide K Tachykinin M 266 P20366 1 RPKPQQFFGLM 11 Substance P Tachykinin 267 P20396 1 QPEAAQQEAVTAAEHPGLDDFLRQVERLLFLRENIQ 218 Pro-thyrotropin- TRH I50 1 RLQGDQGEHSASQIFQSDWLSKRQHPGKREEEEEEG releasing VEEEEEEEGGAVGPHKRQHPGRREDEASWSVDVTQ hormone HKRQHPGRRSPWLAYAVPKRQHPGRRLADPKAQR SWEEEEEEEEREEDLMPEKRQHPGKRALGGPCGPQ GAYGQAGLLLGLLDDLSRSQGAEEKRQHPGRRAA WVREPLEE 268 P20396 1 QHP 3 Thyrotropin- TRH releasing hormone 269 O95399 1 ETPDCFWKYCV 11 Urotensin-2 Urotensin-2 270 Q76510 1 ACFWKYCV 8 Urotensin-2B Urotensin-2 271 P01185 1 CYFQNCPRG 9 Arg-vasopressin Vasopressin/oxytocin 272 P01185 1 ASDRSNATQLDGPAGALLLRLVQLAGAPEPFLPAQP 39 Copeptin Vasopressin/oxytocin A27 1 DAY 273 P01178 1 AAPDLDVRKCLPCGPGGKGRCFGPNICCAEELGCFV 94 Neurophysin 1 Vasopressin/oxytocin GTAEALRCQEENYLPSPCQSGQKACGSGGRCAVLG LCCSPDGCHADPACDAEATFSQR 274 P01185 1 AMSDLELRQCLPCGPGGKGRCFGPSICCADELGCFV 93 Neurophysin 2 Vasopressin/oxytocin GTAEALRCQEENYLPSPCQSGQKACGSGGRCAAFG VCCNDESCVTEPECREGFFIRRA 275 P01178 1 CYIQNCPLG 9 Oxytocin Vasopressin/oxytocin 276 O15240 1 TLQPPSALRRRHYHHALPPSRHYP 24 Antimicrobial VGF peptide VGF[554-577] 277 O15240 1 RPESALLGGSEAGERLLQQGLAQVEA 26 Neuroendocrine VGF regulatory peptide-1 278 O15240 1 QAEATRQAAAQEERLADLASDLLLQYLLQGGARQR 38 Neuroendocrine VGF GLG regulatory peptide-2 279 O15240 1 APPGRPEAQPPPLSSEHKEPVAGDAVPGPKDGSAPE 591 Neurosecretory VGF P1, S12, P202, S205, 5 VRGARNSEPQDEGELFQGVDPRALAAVLLQALDRP protein VGF T477 ASPPAPSGSQQGPEEEAAEALLTETVRSQTHSLPAPE SPEPAAPPRPQTPENGPEASDPSEELEALASLLQELR DFSPSSAKRQQETAAAETETRTHTLTRVNLESPGPE RVWRASWGEFQARVPERAPLPPPAPSQFQARMPDS GPLPETHKFGEGVSSPKTHLGEALAPLSKAYQGVAA PFPKARRPESALLGGSEAGERLLQQGLAQVEAGRR QAEATRQAAAQEERLADLASDLLLQYLLQGGARQR GLGGRGLQEAAEERESAREEEEAEQERRGGEERVG EEDEEAAEAEAEAEEAERARQNALLFAEEEDGEAG AEDKRSQEETPGHRRKEAEGTEEGGEEEDDEEMDP QTIDSLIELSTKLHLPADDVVSIIEEVEEKRKRKKNA PPEPVPPPRAAPAPTHVRSPQPPPPAPAPARDELPDW NEVLPPWDREEDEVYPPGPYHPFPNYIRPRTLQPPSA LRRRHYHHALPPSRHYPGREAQARRAQEEAEAEER RLQEQEELENYIEHVLLRRP
EMBODIMENTS
[0234] 1. An isolated peptide hormone, such as recombinant, such as a neuropeptide comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region.
[0235] 2. The peptide hormone according to embodiment 1, wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with sialic acid capping, such as a structure as illustrated in
[0236] 3. The peptide hormone according to embodiments 1 or 2, wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
[0237] 4. The peptide hormone according to any one of embodiments 1-3, wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
[0238] 5. The peptide hormone according to any one of embodiments 1-4, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
[0239] 6. The peptide hormone according to any one of embodiments 1-5, wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
[0240] 7. The peptide hormone according to any one of embodiments 1-6, wherein the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
[0241] 8. The peptide hormone according to any one of embodiments 1-7, which peptide hormone has improved, such as increased stability and/or circulatory half-life and/or other pharmacokinetic properties, such as improved stability in in vitro assays, plasma and/or bodyfluids.
[0242] 9. The peptide hormone according to any one of embodiments 1-8, which peptide hormone has lower bioactivity in receptor signalling, such as decreased receptor stimulation in in vitro cell assays and/or in man.
[0243] 10. The peptide hormone according to any one of embodiments 1-9, which peptide hormone exhibits improved receptor stimulation in in vitro cell assays and/or in man.
[0244] 11. The peptide hormone according to any one of embodiments 1-10, which peptide hormone exhibits altered blood-brain barrier uptake in animals or in man, such as increased blood-brain barrier uptake in animals or in man, or decreased blood-brain barrier uptake in animals or in human.
[0245] 12. The peptide hormone according to any one of embodiments 1-11, which peptide hormone exhibits receptor sub-type selectivity switch.
[0246] 13. The peptide hormone according to any one of embodiments 1-12, which peptide hormone is specific to one or more tissue in human, such as specific to tissue of the nervous system.
[0247] 14. The peptide hormone according to any one of embodiments 1-13, which peptide hormone is selected from any one of tables 4, 5, or 6, such as selected from the list consisting of a peptide of the Neuropeptide Y family, such as NPY, PPY and PYY; a peptide of the Glucagon/Secretin family, such as GIP, Glucagon, GLP-1, GLP-2, PACAP, Secretin, PHM-27/PHV-42, Somatoliberin, and VIP; a peptide of the Natriuretic peptide family, such as ANP, BNP and CNP, a peptide of the calcitonin family, such as calcitonin, and amylin.
[0248] 15. The peptide hormone according to any one of embodiments 1-14, which peptide hormone is not found in nature.
[0249] 16. The peptide hormone according to any one of embodiments 1-15, which peptide hormone is a truncated version or a variant as compared to the corresponding wild-type peptide hormone found in nature.
[0250] 17. The peptide hormone according to any one of embodiments 1-16, which peptide hormone is selected from any one of table 6 comprising one or more O-linked glycan at a site as indicated in table 6, such as at a bold underlined position and/or an italic underlined position.
[0251] 18. The peptide hormone according to any one of embodiments 1-17, which peptide hormone is selected from any one of table 6 comprising at least, not more than, or the exact number of O-linked glycan sites as indicated in table 6.
[0252] 19. The peptide hormone according to any one of embodiments 1-16, which peptide hormone is selected from any one of table 5.
[0253] 20. A host cell comprising two or more glycosyltransferase genes that have been inactivated such that
(a) Homogenous Tn (GalNAc) glycosylation is obtained by inactivation and/or downregulation of one or more genes selected from COSMC and C1GALT1;
(b) Homogenous T (Gal/GalNAc) glycosylation is obtained by inactivation and/or downregulation of one or more genes selected from GCNT1, GCNT3, GCNT4, B3GNT6; and
(c) Homogenous ST or STn glycosylation is obtained by inactivation and/or downregulation of one or more genes selected from ST6GALNAC1-6, ST3GAL1, GCNT3, GCNT4, B3GNT6.
[0254] 21. The host cell according to embodiment 20, further comprising a gene encoding an exogenous peptide hormone, such as a peptide hormone as defined in any one of embodiments 14-19.
[0255] 22. A method for producing an isolated peptide hormone comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region, the method comprising; [0256] a) inactivation and/or downregulation of one or more glycosyltransferases, and/or knock in of one or more glycosyltransferases, or any combination hereof in a host cell, and [0257] b) expression of said peptide hormone in said host cell.
[0258] 23. The method according to embodiment 22, wherein one or more genes selected from COSMC, C1GALT1, GCNT1, GCNT3, GCNT4, B3GNT6, ST6GALNAC1-6, ST3GAL1 has been inactivated and/or downregulated.
[0259] 24. The method according to embodiments 22 or 23, wherein said peptide hormone produced is as defined in any one of embodiments 1-19.
[0260] 25. The method according to any one of embodiments 22-24, wherein said host cell is as defined in any of embodiments 20-21.
[0261] 26. A method for the production of recombinant glycosylated peptide hormones that do not have specific types of glycosylation, the method comprising the step of inactivating two or more glycogenes to block and/or truncate one or more glycosylation pathways.
[0262] 27. A method for the production of an isolated peptide hormone, such as a neuropeptide comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region, said method comprising [0263] a) providing a non-O-glycosylated peptide hormone; and [0264] b) treating said non-O-glycosylated synthetic peptide hormone with one or more recombinant purified glycosyl transferase, such as a GalNAc-transferase, such as GalNAc-T1, T2, T3, T4, T5, T6, T7, T10, T11, T12, T13, T14, and/or T16, and/or a Galactosyl-transferases (C1GalT1) and/or a sialyl-transferases, such as ST6GalNAc1 and/or ST3Gall under conditions to add one or more specific O-linked glycan to said peptide hormone.
[0265] 28. The method according to embodiment 27, wherein said non-O-glycosylated peptide hormone is provided as a chemically produced peptide hormone produced using solid phase peptide synthesis Fmoc SPPS.
[0266] 29. The method according to embodiment 27, wherein said non-O-glycosylated peptide hormone is provided as a recombinantly produced peptide hormone, such as produced in a production cell line.
[0267] 30. The method according to any one of embodiments 27-29, wherein said peptide hormone produced is as defined in any one of embodiments 1-19.
[0268] 31. A method for the production of an isolated peptide hormone, such as a neuropeptide comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region, said method comprising the building of said peptide hormone using solid phase peptide synthesis Fmoc SPPS including the use of glycosylated amino acids building blocks at said predetermined specific site(s).
[0269] 32. The method according to embodiment 31, wherein the peptide hormone produced is as defined in any one of embodiments 1-19.
ADDITIONAL EMBODIMENTS
[0270] a. An isolated peptide hormone, such as recombinant, such as a neuropeptide comprising one or more O-linked glycan at a predetermined specific site, such as in the receptor-binding region.
[0271] b. The peptide hormone according to embodiment a), wherein the one or more O-glycan structures include a glycan structure selected from a core1, core2, core3, or core4 structure with sialic acid capping, such as a structure as illustrated in
[0272] c. The peptide hormone according to embodiments a) or b), wherein the one or more O-glycan structures include a Tn (GalNAc) structure.
[0273] d. The peptide hormone according to any one of embodiments a) to c), wherein the one or more O-glycan structures include Tn (GalNAc) structure with one sialic acid capping (alpha2-6).
[0274] e. The peptide hormone according to any one of embodiments a)-d), wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha2-6).
[0275] f. The peptide hormone according to any one of embodiments a)-e), wherein the one or more O-glycan structures include the core1 structures with one sialic acid capping (alpha 2-3)
[0276] g. The peptide hormone according to any one of embodiments a)-f), wherein the one or more O-glycan structures include the core1 structures with two sialic acids capping (alpha 2-3 and alpha 2-6).
[0277] h. The peptide hormone according to any one of embodiments a)-g), which peptide hormone has improved, such as increased stability and/or circulatory half-life and/or other pharmacokinetic properties, such as improved stability in in vitro assays, plasma and/or bodyfluids.
[0278] i. The peptide hormone according to any one of embodiments a)-h), which peptide hormone has lower bioactivity in receptor signalling, such as decreased receptor stimulation in in vitro cell assays and/or in man.
[0279] j. The peptide hormone according to any one of embodiments a)-i), which peptide hormone exhibits improved receptor stimulation in in vitro cell assays and/or in man.
[0280] k. The peptide hormone according to any one of embodiments a)-j), which peptide hormone exhibits altered blood-brain barrier uptake in animals or in man, such as increased blood-brain barrier uptake in animals or in man, or decreased blood-brain barrier uptake in animals or in human.
[0281] l. The peptide hormone according to any one of embodiments a)-k), which peptide hormone exhibits receptor sub-type selectivity switch.
[0282] m. The peptide hormone according to any one of embodiments a)-l), which peptide hormone is selected from any one of tables 4, 5, or 6, such as selected from the list consisting of a peptide of the Neuropeptide Y family, such as NPY, PPY and PYY; a peptide of the Glucagon/Secretin family, such as GIP, Glucagon, GLP-1, GLP-2, PACAP, Secretin, PHM-27/PHV-42, Somatoliberin, and VIP; a peptide of the Natriuretic peptide family, such as ANP, BNP and CNP, a peptide of the calcitonin family, such as calcitonin, and amylin.
[0283] n. The peptide hormone according to any one of embodiments a)-m), which peptide hormone is not found in nature, such as a truncated version or a variant as compared to the corresponding wild-type peptide hormone found in nature.
[0284] o. The peptide hormone according to any one of embodiments a)-n), which peptide hormone is selected from any one of table 6 comprising one or more O-linked glycan at a site as indicated in table 6, such as at a bold underlined position and/or an italic underlined position.
LIST OF REFERENCES
[0285] 1 Seidah, N. G. What lies ahead for the proprotein convertases? Ann N Y Acad Sci 1220, 149-161, doi: 10.1111/j. 1749-6632.2010.05883.x (2011). [0286] 2 Milgram, S. L., Mains, R. E. & Eipper, B. A. COOH-terminal signals mediate the trafficking of a peptide processing enzyme in endocrine cells. J Cell Biol 121, 23-36 (1993). [0287] 3 Bundgaard, J. R., Vuust, J. & Rehfeld, J. F. Tyrosine O-sulfation promotes proteolytic processing of progastrin. The EMBO journal 14, 3073-3079 (1995). [0288] 4 Drazic, A., Myklebust, L. M., Ree, R. & Arnesen, T. The world of protein acetylation. Biochim Biophys Acta 1864, 1372-1401, doi:10.1016/j.bbapap.2016.06.007 (2016). [0289] 5 Secher, A. et at. Analytic framework for peptidomics applied to large-scale neuropeptide identification. Nature communications 7, 11436, doi:10.1038/ncomms11436 (2016). [0290] 6 Mentlein, R. Dipeptidyl-peptidase IV (CD26)—role in the inactivation of regulatory peptides. Regulatory peptides 85, 9-24 (1999). [0291] 7 Potter, L. R. Natriuretic peptide metabolism, clearance and degradation. Febs J 278, 1808-1817, doi:10.1111/j.1742-4658.2011.08082.x (2011). [0292] 8 Deacon, C. F., Nauck, M. A., Meier, J., Hucking, K. & Holst, J. J. Degradation of endogenous and exogenous gastric inhibitory polypeptide in healthy and in type 2 diabetic subjects as revealed using a new assay for the intact peptide. J Clin Endocrinol Metab 85, 3575-3581, doi: 10.1210/jcem.85.10.6855 (2000). [0293] 9 Halim, A., Ruetschi, U., Larson, G. & Nilsson, J. LC-MS/MS characterization of O-glycosylation sites and glycan structures of human cerebrospinal fluid glycoproteins. Journal of proteome research 12, 573-584, doi:10.1021/pr300963 h (2013). [0294] 10 Halim, A., Nilsson, J., Ruetschi, U., Hesse, C. & Larson, G. Human urinary glycoproteomics; attachment site specific analysis of N- and O-linked glycosylations by CID and ECD. Molecular & cellular proteomics: MCP 11, Mill 013649, doi: 10.1074/mcp.M 111.013649 (2012). [0295] 11 Khetarpal, S. A. et at Loss of Function of GALNT2 Lowers High-Density Lipoproteins in Humans, Nonhuman Primates, and Rodents. Cell Metab 24, 234-245, doi: 10.1016/j.cmet.2016.07.012 (2016). [0296] 12 King, S. L. et at Characterizing the O-glycosylation landscape of human plasma, platelets, and endothelial cells. Blood Advances 1, 429-442, doi: 10.1182/bloodadvances. 2016002121 (2017). [0297] 13 Yu, Q. et at Targeted Mass Spectrometry Approach Enabled Discovery of O-Glycosylated Insulin and Related Signaling Peptides in Mouse and Human Pancreatic Islets. Anal Chem, doi:10.1021/acs.analchem.7b01926 (2017). [0298] 14 Cao, F. et at Detection of Biosynthetic Precursors, Discovery of Glycosylated Forms, and Homeostasis of Calcitonin in Human Cancer Cells. Anal Chem 89, 6992-6999, doi: 10.1021/acs.analchem.7b00457 (2017). [0299] 15 Bennett, E. P. et at Control of mucin-type O-glycosylation: A classification of the polypeptide GalNAc-transferase gene family. Glycobiology 22, 736-756, doi:Doi 10.1093/Glycob/Cwr182 (2012). [0300] 16 Topaz, O. et at Mutations in GALNT3, encoding a protein involved in O-linked glycosylation, cause familial tumoral calcinosis. Nat Genet 36, 579-581, doi:10.1038/ng1358 (2004). [0301] 17 Kato, K. et at Polypeptide GalNAc-transferase T3 and familial tumoral calcinosis. Secretion of fibroblast growth factor 23 requires O-glycosylation. J Biol Chem 281, 18370-18377, doi:M602469200 [pii]10.1074/jbe.M602469200 (2006). [0302] 18 Schjoldager, K. T. et at Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells. Proc Natl Acad Sci U SA 109, 9893-9898, doi: 10.1073/pnas. 1203563109 (2012). [0303] 19 Chretien, M. & Mbikay, M. 60 YEARS OF POMC: From the prohormone theory to pro-opiomelanocortin and to proprotein convertases (PCSK1 to PCSK9). J Mol Endocrinol 56, T49-62, doi: 10.1530/JME-15-0261 (2016). [0304] 20 Walsh, G. & Jefferis, R. Post-translational modifications in the context of therapeutic proteins. Nature biotechnology 24, 1241-1252, doi:10.1038/nbt1252 (2006). [0305] 21 Tenno, M. et al. Initiation of protein O glycosylation by the polypeptide GalNAcT-1 in vascular biology and humoral immunity. Mol Cell Biol 27, 8783-8796, doi:10.1128/MCB.01204-07 (2007). [0306] 22 Tian, E. & Ten Hagen, K. G. A UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase is required for epithelial tube formation. J Biol Chem 282, 606-614, doi: 10.1074/jbe.M606268200 (2007). [0307] 23 Freire-de-Lima, L. et al. Involvement of O-glycosylation defining oncofetal fibronectin in epithelial-mesenchymal transition process. Proc Natl Acad Sci USA 108, 17690-17695, doi: 10.1073/pnas. 1115191108 (2011). [0308] 24 Schjoldager, K. T. & Clausen, H. Site-specific protein O-glycosylation modulates proprotein processing—deciphering specific functions of the large polypeptide GalNAc-transferase gene family. Biochim Biophys Acta 1820, 2079-2094, doi:10.1016/j.bbagen.2012.09.014 (2012). [0309] 25 Tagliabracci, V. S. et al. Dynamic regulation of FGF23 by Fam20C phosphorylation, GalNAc-T3 glycosylation, and furin proteolysis. Proc Natl Acad Sci USA 111, 5520-5525, doi: 10.1073/pnas. 1402218111 (2014). [0310] 26 Goth, C. K. et at. A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. Proc Natl Acad Sci USA 112, 14623-14628, doi: 10.1073/pnas. 1511175112 (2015). [0311] 27 Norden, R. et al. O-linked glycosylation of the mucin domain of the herpes simplex virus type 1-specific glycoprotein gC-1 is temporally regulated in a seed-and-spread manner. The Journal of biological chemistry 290, 5078-5091, doi:10.1074/jbc.M114.616409 (2015). [0312] 28 Schjoldager, K. T. et al. Deconstruction of O-glycosylation—GalNAc-T isoforms direct distinct subsets of the O-glycoproteome. EMBO Rep 16, 1713-1722, doi: 10.15252/embr.201540796 (2015). [0313] 29 Tian, E. et al. Galnt1 is required for normal heart valve development and cardiac function. PLoS One 10, e0115861, doi:10.1371/journal.pone.0115861 (2015). [0314] 30 Akasaka-Manya, K. et al. Excess APP O-glycosylation by GalNAc-T6 decreases Abeta production. Journal of biochemistry 161, 99-111, doi: 10.1093/jb/mvw056 (2017). [0315] 31 Lin, J. et al. GALNT6 Stabilizes GRP78 Protein by O-glycosylation and Enhances its Activity to Suppress Apoptosis Under Stress Condition. Neoplasia (New York, N.Y. 19, 43-53, doi: 10.1016/j.neo.2016.11.007 (2017). [0316] 32 Goth, C. K. et al. Site-specific O-glycosylation by Polypeptide GalNAc-transferase T2 Co-regulates Beta1-adrenergic Receptor N-terminal Cleavage. J Biol Chem, doi: 10.1074/j be. Ml 16.730614 (2017). [0317] 33 Schjoldager, K. T. B. G. et al. A Systematic Study of Site-specific GalNAc-type O-Glycosylation Modulating Proprotein Convertase Processing. Journal of Biological Chemistry 286, 40122-40132, doi:10.1074/jbc.M111.287912 (2011). [0318] 34 Schjoldager, K. T. B. G. et al. O-Glycosylation Modulates Proprotein Convertase Activation of Angiopoietin-like Protein 3 POSSIBLE ROLE OF POLYPEPTIDE GalNAc-TRANSFERASE-2 IN REGULATION OF CONCENTRATIONS OF PLASMA LIPIDS. Journal of Biological Chemistry 285, 36293-36303, doi:10.1074/jbc.M110.156950 (2010). [0319] 35 Pedersen, N. B. et al. Low density lipoprotein receptor class a repeats are o-glycosylated in linker regions. J Biol Chem 289, 17312-17324, doi:10.1074/jbc.M113.545053 (2014). [0320] 36 Wang, S. et al. Site-specific O-glycosylation of members of the low-density lipoprotein receptor superfamily enhances ligand interactions. J Biol Chem, doi: 10.1074/j be. Ml 17.817981 (2018). [0321] 37 Pedragosa-Badia, X., Stichel, J. & Beck-Sickinger, A. G. Neuropeptide Y receptors: how to get subtype selectivity. Front Endocrinol (Lausanne) 4, 5, doi: 10.3389/fendo.2013.00005 (2013). [0322] 38 Tan, T. et al. The effect of a subcutaneous infusion of GLP-1, OXM and PYY on Energy intake and Expenditure in Obese volunteers. J Clin Endocrinol Metab, doi:10.1210/jc.2017-00469 (2017). [0323] 39 van Deursen, V. M. et al. Nesiritide, renal function, and associated outcomes during hospitalization for acute decompensated heart failure: results from the Acute Study of Clinical Effectiveness of Nesiritide and Decompensated Heart Failure (ASCEND-HF). Circulation 130, 958-965, doi:10.1161/CIRCULATIONAHA.113.003046 (2014). [0324] 40 Boerrigter, G. & Burnett, J. C., Jr. Natriuretic peptides renal protective after all? J Am Coll Cardiol 58, 904-906, doi:10.1016/j.jacc.2010.12.053 (2011). [0325] 41 Hata, N. et al. Effects of carperitide on the long-term prognosis of patients with acute decompensated chronic heart failure: the PROTECT multicenter randomized controlled study. Circ J 72, 1787-1793 (2008). [0326] 42 Fosgerau, K. & Hoffmann, T. Peptide therapeutics: current status and future directions. Drug Discov Today 20, 122-128, doi:10.1016/j.drudis.2014.10.003 (2015). [0327] 43 Yi, J., Warunek, D. & Craft, D. Degradation and Stabilization of Peptide Hormones in Human Blood Specimens. PLoS One 10, e0134427, doi: 10.1371/journal.pone.0134427 (2015). [0328] 44 Chapter, M. C. et at. Chemical modification of class II G protein-coupled receptor ligands: frontiers in the development of peptide analogs as neuroendocrine pharmacological therapies. Pharmacol Ther 125, 39-54, doi:10.1016/j.pharmthera.2009.07.006 (2010). [0329] 45 DeFrees, S. et al. GlycoPEGylation of recombinant therapeutic proteins produced in Escherichia coli. Glycoblology 16, 833-843, doi:10.1093/glycob/cwl004 (2006). [0330] 46 Meurer, J. A., Colca, J. R., Burton, P. S. & Elhammer, A. P. Properties of native and in vitro glycosylated forms of the glucagon-like peptide-1 receptor antagonist exendin(9-39). Metabolism: clinical and experimental 48, 716-724 (1999). [0331] 47 Egleton, R. D. et al. Improved blood-brain barrier penetration and enhanced analgesia of an opioid peptide by glycosylation. J Pharmacol Exp Ther 299, 967-972 (2001). [0332] 48 Kihlberg, J. et al. Glycosylated peptide hormones: pharmacological properties and conformational studies of analogues of [1-desamino,8-D-arginine]vasopressin. J Med Chem 38, 161-169 (1995). [0333] 49 Ueda, T. et al. Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1: effect of glycosylation on proteolytic resistance and in vivo blood glucose-lowering activity. Journal of the American Chemical Society 131, 6237-6245, doi:10.1021/ja900261 g (2009). [0334] 50 Pedersen, S. L., Steentoft, C., Vrang, N. & Jensen, K. J. Glyco-scan: varying glycosylation in the sequence of the peptide hormone PYY3-36 and its effect on receptor selectivity. Chembiochem 11, 366-374, doi:10.1002/cbic.200900661 (2010). [0335] 51 Sato, M. et al. Glycoinsulins: dendritic sialyloligosaccharide-displaying insulins showing a prolonged blood-sugar-lowering activity. Journal of the American Chemical Society 126, 14013-14022, doi: 10.1021/ja046426l (2004). [0336] 52 Dangoor, D. et al. Novel glycosylated VIP analogs: synthesis, biological activity, and metabolic stability. J Pept Sci 14, 321-328, doi: 10.1002/psc.932 (2008). [0337] 53 Semenov, A. G. et al. Processing of pro-brain natriuretic peptide is suppressed by O-glycosylation in the region close to the cleavage site. Clin Chem 55, 489-498, doi: 10.1373/clinchem.2008.113373 (2009). [0338] 54 Duguay, S. J. et al. Post-translational processing of the insulin-like growth factor-2 precursor. Analysis of O-glycosylation and endoproteolysis. The Journal of biological chemistry 273, 18443-18451 (1998). [0339] 55 Birch, N. P., Estivariz, F. E., Bennett, H. P. & Loh, Y. P. Differential glycosylation of N-POMC1-77 regulates the production of gamma 3-MSH by purified pro-opiomelanocortin converting enzyme. A possible mechanism for tissue-specific processing. FEBS Lett 290, 191-194 (1991). [0340] 56 Patzelt, C. & Weber, B. Early O-glycosidic glycosylation of proglucagon in pancreatic islets: an unusual type of prohormonal modification. The EMBO journal 5, 2103-2108 (1986). [0341] 57 Luckenbill, K. N. et al. Cross-reactivity of BNP, NT-proBNP, and proBNP in commercial BNP and NT-proBNP assays: preliminary observations from the IFCC Committee for Standardization of Markers of Cardiac Damage. Clin Chem 54, 619-621, doi: 10.1373/clinchem.2007.097998 (2008). [0342] 58 Stovold, R. et al. Biomarkers for small cell lung cancer: neuroendocrine, epithelial and circulating tumour cells. Lung Cancer 76, 263-268, doi:10.1016/j.lungcan.2011.11.015 (2012). [0343] 59 Wang, J. et al. Identification of kininogen-1 as a serum biomarker for the early detection of advanced colorectal adenoma and colorectal cancer. PLoS One 8, e70519, doi:10.1371/journal.pone.0070519 (2013). [0344] 60 Goetze, J. P., Alehagen, U., Flyvbjerg, A. & Rehfeld, J. F. Chromogranin A as a biomarker in cardiovascular disease. Biomark Med 8, 133-140, doi:10.2217/bmm.13.102 (2014). [0345] 61 Nielsen, S. J., Iversen, P., Rehfeld, J. F., Jensen, H. L. & Goetze, J. P. C-type natriuretic peptide in prostate cancer. APMIS 117, 60-67, doi: 10.1111/j. 1600-0463.2008.00016.x 60 (2009). [0346] 62 Ju, T., Aryal, R. P., Stowell, C. J. & Cummings, R. D. Regulation of protein O-glycosylation by the endoplasmic reticulum-localized molecular chaperone Cosmc. J Cell Biol 182, 531-542, doi:10.1083/jcb.200711151 (2008). [0347] 63 Steentoft, C. et al. Mining the O-glycoproteome using zinc-finger nuclease-glycoengineered SimpleCell lines. Nat Methods 8, 977-982, doi:10.1038/nmeth.1731 (2011). [0348] 64 Steentoft, C. et al. Precision genome editing: A small revolution for glycobiology. Glycobiology 24, 663-680, doi: 10.1093/glycob/cwu046 (2014). [0349] 65 Yamane-Ohnuki, N. et at Establishment of FUT8 knockout Chinese hamster ovary cells: an ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity. Biotechnol Bioeng 87, 614-622, doi: 10.1002/bit. 20151 (2004). [0350] 66 Malphettes, L. et at Highly efficient deletion of FUT8 in CHO cell lines using zinc-finger nucleases yields cells that produce completely nonfucosylated antibodies. Biotechnol Bioeng 106, 774-783, doi: 10.1002/bit.22751 (2010). [0351] 67 Ronda, C. et at Accelerating genome editing in CHO cells using CRISPR Cas9 and CRISPy, a web-based target finding tool. Biotechnol Bioeng 111, 1604-1616, doi: 10.1002/bit.25233 (2014). [0352] 68 El Mai, N., Donadio-Andrei, S., Iss, C., Calabro, V. & Ronin, C. Engineering a human-like glycosylation to produce therapeutic glycoproteins based on 6-linked sialylation in CHO cells. Methods in molecular biology 988, 19-29, doi:10.1007/978-1-62703-327-5_2 (2013). [0353] 69 Sewell, R. et at The ST6GalNAc-I sialyltransferase localizes throughout the Golgi and is responsible for the synthesis of the tumor-associated sialyl-Tn O-glycan in human breast cancer. The Journal of biological chemistry 281, 3586-3594, doi: 10.1074/jbc.M511826200 (2006). [0354] 70 Torang, S. et at In vivo and in vitro degradation of peptide YY3-36 to inactive peptide YY3-34 in humans. Am J Physiol Regul Integr Comp Physiol 310, R866-874, doi: 10.1152/ajpregu.00394.2015 (2016). [0355] 71 Steentoft, C. et at Precision mapping of the human O-GalNAc glycoproteome through SimpleCell technology. The EMBO journal 32, 1478-1488, doi:10.1038/emboj.2013.79 (2013). [0356] 72 Levery, S. B. et at Advances in mass spectrometry driven O-glycoproteomics. Biochim Biophys Acta, doi:10.1016/j.bbagen.2014.09.026 (2014). [0357] 73 Schietinger, A. et at A mutant chaperone converts a wild-type protein into a tumor-specific antigen. Science 314, 304-308, doi:10.1126/science.1129200 (2006). [0358] 74 Yabu, M., Korekane, H. & Miyamoto, Y. Precise structural analysis of O-linked oligosaccharides in human serum. Glycobiology 24, 542-553, doi:10.1093/glycob/cwu022 (2014). [0359] 75 Locke, A. E. et at Genetic studies of body mass index yield new insights for obesity biology. Nature 518, 197-206, doi:10.1038/nature14177 (2015). [0360] 76 Zougman, A. et at Integrated analysis of the cerebrospinal fluid peptidome and proteome. Journal of proteome research 7, 386-399, doi:10.1021/pr070501k (2008). [0361] 77 Lottspeich, F., Kellermann, J., Henschen, A., Foertsch, B. & Muller-Esterl, W. The amino acid sequence of the light chain of human high-molecular-mass kininogen. European journal of biochemistry/FEBS 152, 307-314 (1985). [0362] 78 Makimattila, S., Fineman, M. S. & Yki-Jarvinen, H. Deficiency of total and nonglycosylated amylin in plasma characterizes subjects with impaired glucose tolerance and type 2 diabetes. J Clin Endocrinol Metab 85, 2822-2827, doi: 10.1210/jcem.85.8.6721 (2000). [0363] 79 Schellenberger, U. et at The precursor to B-type natriuretic peptide is an O-linked glycoprotein. Arch Biochem Biophys 451, 160-166, doi:10.1016/j.abb.2006.03.028 (2006). [0364] 80 Pal, K., Melcher, K. & Xu, H. E. Structure and mechanism for recognition of peptide hormones by Class B G-protein-coupled receptors. Acta Pharmacol Sin 33, 300-311, 55 doi:10.1038/aps.2011.170 (2012). [0365] 81 Naot, D. & Cornish, J. The role of peptides and receptors of the calcitonin family in the regulation of bone metabolism. Bone 43, 813-818, doi: 10.1016/j.bone.2008.07.003 (2008). [0366] 82 Muff, R., Born, W. & Fischer, J. A. Calcitonin, calcitonin gene-related peptide, adrenomedullin and amylin: homologous peptides, separate receptors and overlapping biological actions. European journal of endocrinology/European Federation of Endocrine Societies 133, 17-20 (1995). [0367] 83 Tagashira, M., Iijima, H. & Toma, K. An NMR study of O-glycosylation induced structural changes in the alpha-helix of calcitonin. Glycoconj J 19, 43-52 (2002). [0368] 84 Fisone, G. et al. N-terminal galanin-(1-16) fragment is an agonist at the hippocampal galanin receptor. Proc Natl Acad Sci USA 86, 9588-9591 (1989). [0369] 85 Cabrele, C. & Beck-Sickinger, A. G. Molecular characterization of the ligand-receptor interaction of the neuropeptide Y family. J Pept Sci 6, 97-122, doi: 10.1002/(SICI)1099-1387(200003)6:3<97::AID-PSC236>3.0.CO;2-E (2000). [0370] 86 Nicole, P. et al. Identification of key residues for interaction of vasoactive intestinal peptide with human VPAC1 and VPAC2 receptors and development of a highly selective VPAC1 receptor agonist. Alanine scanning and molecular modeling of the peptide. J Biol Chem 275, 24003-24012, doi: 10.1074/jbc.M002325200 (2000). [0371] 87 Cerda-Reverter, J. M. & Larhammar, D. Neuropeptide Y family of peptides: structure, anatomical expression, function, and molecular evolution. Biochem Cell Biol 78, 371-392 (2000). [0372] 88 Scarborough, R. M. et al. D-amino acid-substituted atrial natriuretic peptide analogs reveal novel receptor recognition requirements. J Biol Chem 263, 16818-16822 (1988). [0373] 89 Campbell, D. J. Long-term neprilysin inhibition—implications for ARNIs. Nat Rev Cardiol 14, 171-186, doi:10.1038/nrcardio.2016.200 (2017). [0374] 90 Lambeir, A. M., Durinx, C., Scharpe, S. & De Meester, I. Dipeptidyl-peptidase IV from bench to bedside: an update on structural properties, functions, and clinical aspects of the enzyme DPP IV. Crit Rev Clin Lab Sci 40, 209-294, doi:10.1080/713609354 (2003). [0375] 91 Nauck, M. Incretin therapies: highlighting common features and differences in the modes of action of glucagon-like peptide-1 receptor agonists and dipeptidyl peptidase-4 inhibitors. Diabetes Obes Metab 18, 203-216, doi: 10.1111/dom. 12591 (2016). [0376] 92 Deacon, C. F. et al. Dipeptidyl peptidase IV resistant analogues of glucagon-like peptide-1 which have extended metabolic stability and improved biological activity. Diabetologia 41, 271-278, doi: 10.1007/s001250050903 (1998). [0377] 93 Plamboeck, A., Holst, J. J., Carr, R. D. & Deacon, C. F. Neutral endopeptidase 24.11 and dipeptidyl peptidase IV are both mediators of the degradation of glucagon-like peptide 1 in the anaesthetised pig. Diabetologia 48, 1882-1890, doi:10.1007/s00125-005-1847-7 (2005). [0378] 94 Hupe-Sodmann, K. et al. Characterisation of the processing by human neutral endopeptidase 24.11 of GLP-1(7-36) amide and comparison of the substrate specificity of the enzyme for other glucagon-like peptides. Regulatory peptides 58, 149-156 (1995). [0379] 95 Torang, S., Veedfald, S., Rosenkilde, M. M., Hartmann, B. & Holst, J. J. The anorexic hormone Peptide YY3-36 is rapidly metabolized to inactive Peptide YY3-34 in vivo. Physiol Rep 3, doi: 10.14814/phy2.12455 (2015).