Provided herein are novel compounds, compositions, that can used as clickable, photo-affinity labeling (PAL) probes methods for use in screening NAD+ binding proteins.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P).sup.+) or derivative thereof; an NAD(P).sup.+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

High mitochondrial ATP is a metabolic trait that confers hyper-proliferation, sternness, anchorage-independence, anti-oxidant capacity and multi-drug resistance in cancer cells. Under the present approach, intracellular ATP levels may be used as a metabolic biomarker to identify, separate, and purify an aggressive and hyper-proliferative cancer stem cell (“CSC”) phenotype. Further, ATP may be combined with other CSC markers, e.g., CD44 or ALDH-activity, to beneficially fractionate the CSC population into sub-populations. For example, ATP-high/ CD44-high CSC sub-populations showed twice the level of anchorage-independent growth compared to ATP-low/CD44-high CSC sub-populations. Also disclosed are complementary bioinformatic data that implicate mitochondrial ATP synthesis in stemness, metastasis, and the detection of circulating tumor cells (“CTCs”), and a five-member, ATP-related metastasis gene-signature (ABCA2, ATP5F1C, COX20, NDUFA2 and UQCRB). The gene signature of the present approach may be used to identify CSCs having a dramatic increase in cell migration and invasion in vitro capacity, as well as spontaneous metastasis in vivo. The present approach also provides a cellular platform for systematically targeting sternness, multi-drug resistance, and metastasis in cancer cells.

A method for detecting an analyte is described in which the simultaneously binding of two fusion proteins (i.e., a sandwich assay in solution) is used, bringing two halves of a split enzyme together to produce product, which is detected via a FRET-based biosensor. The method may incorporate an autocatalytic feedback loop that responds to enzymatic product by producing more product to provide ultrasensitive, bistable detection of analyte that is tunable over several orders of magnitude. This system is broadly applicable for protein and small molecule detection.

The invention relates to antibodies or antibody fragments capable of binding to G-protein alpha, nucleic acid sequences coding for the antibody, vectors comprising the nucleic acid sequence, cells comprising the vector or the nucleic acid sequence and a kit-of-parts comprising (i) the antibody or antibody fragment or the composition and (ii) a source of GTP labeled with a member of a pair of RET partners.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P).sup.+) or derivative thereof; an NAD(P).sup.+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

This invention pertains to a fluorescence lifetime imaging microscopy (FLIM)-based method for the quantitative three-dimensional (3D) imaging of the metabolic status of a lesion, including the acquisition of FLIM data of a least one anaerobic glycolysis marker. This method applies in particular to lesions such as wounds and tumours, and to markers such as the NADH/NAD+ ratio, the NAD(P)H/NAD+ ratio, the NADH/FAD ratio, the NAD(P)H/FAD ratio or the FADH/FAD ratio as a first marker, and their optional combination with a second marker such as pH, oxygen tension, glucose, pyruvate and lactate.

The present invention provides a method to identify compounds useful in inhibiting the adverse effects of excessive FABP4 on the modulation of NDPK-ADK agonism of G protein-coupled receptors (GPCR) and channels in FABP4-mediated disorders. It has been surprisingly discovered that the fatty acid binding protein 4 (FABP4) inhibits the ability of the nucleoside diphosphate kinase (NDPK) and adenosine kinase (ADK) complex to agonize GPCRs on target cells by forming an NDPK-ADK/FABP4 complex, resulting in, amongst other things, impaired or reduced insulin secretion in islet β-cells and an increase in glucose levels in the bloodstream. By inhibiting the formation of the NDPK-ADK/FABP4 complex, or inhibiting FABP4 downregulation of NDPK-ADK complex modulation of GPCRs, it has been discovered that FABP4-medited effects can be blunted, including the modulation of islet β-cell insulin secretion, providing for a reduction in glucose levels and the attenuation of metabolic dysfunction.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P).sup.+) or derivative thereof; an NAD(P).sup.+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention also relates to compounds, which are useful for intra-molecular fluorescence resonance energy transfer (FRET), comprising the oxidized form of a carbaNADH-based first fluorophore and a second fluorophore that is excitable at a wave-length of between 445 to 540 nm and that has an emission maximum of greater than 560 nm, and methods, kits and compositions related thereto.