This invention relates to methods of identifying, synthesizing, optimizing and profiling compounds that are inhibitors or activators of proteins, both naturally occurring endogenous proteins as well as certain variant forms of endogenous proteins, and novel methods of identifying such variants. The method accelerates the identification and development of compounds as potential therapeutically effective drugs by simplifying the pharmaceutical discovery and creation process through improvements in hit identification, lead optimization, biological profiling, and rapid elimination of toxic compounds. Implementation results in overall cost reductions in the drug discovery process resulting from the corresponding increases in efficiency.

Disclosed are methods of determining activity of CDK4 and CDK6 variants upon exposure to CDK inhibitors, methods for determining activity of a Rb variant, methods for determining the activity of a p16 variant in a cell, and methods for determining the sensitivity of a CDK4 variant or a CDK6 variant to p16 in a cell. Stable cell lines for determining activity of CDK4 variants, CDK6 variants, Rb variants, and p16 variants are also disclosed.

The disclosure provides polypeptides comprising an engineered p27, or a fragment thereof. Such polypeptides may be used to form trimeric protein complexes with a cyclin-dependent kinase 4 (Cdk4) (or a variant thereof) or Cdk6 (or a variant thereof), and a cyclin D (CycD) or a variant thereof.

The present invention relates to enzyme inhibitors. More specifically, the present invention relates to ligand-directed covalent modification of proteins; method of designing same; pharmaceutical formulation of same; and method of use.

Described are methods for developing treatment plans for patients with cancer and other diseases based on drug binding interactions of kinase inhibitors. Also described are methods for determining the mutational status of a kinase and kinase occupancy.

A method, a composition and a kit for screening an ALK or ROS-1 kinase inhibitor are disclosed in the present specification. In an aspect, by the method, composition and kit for screening an ALK or ROS-1 kinase inhibitor according to the present disclosure, it is possible to conduct simultaneous quantitative and qualitative analysis and faster screening of a larger number of candidate substances as compared with conventional molecular biological experimental methods and to grasp the overall level of change in a plurality of different metabolites in a cell line by candidate substances of ALK or ROS-1 kinase inhibitor and thus the screening efficiency for drugs which exert an ALK or ROS-1 kinase inhibitory effect is excellent. Consequently, the present disclosure has an advantage of being able to be used in various ways in the development of new drugs which exert an ALK or ROS-1 kinase inhibitory effect.

The present invention provides a method for controlling a speed of a polypeptide passing through a nanopore and use thereof in determining an amino acid sequence of a polypeptide. Specifically, the method comprises: conjugating a polynucleotide to the polypeptide to give a polynucleotide-polypeptide conjugate, and applying a voltage across the nanopore in the presence of a polynucleotide binding enzyme to move the conjugate through the nanopore. The polynucleotide binding enzyme controls the movement of the polynucleotide and thereby controls the movement of the conjugated polypeptide in the nanopore, thus controlling the speed of the polypeptide passing through the nanopore. While controlling the speed of the polypeptide, the present invention reads a nanopore current signal during the process of the polypeptide passing through the nanopore to give an electrical signal of the polypeptide. The electrical signal can be further used to acquire an amino acid sequence of the polypeptide, to identify the polypeptide or a part thereof, or to establish a library of polypeptide electrical signals.

The present invention includes method and system for detecting kinase activity in vivo, comprising: providing a cell that comprises one or more mutated kinases, wherein the one or more mutated kinases comprise a mutation that enlarges an ATP binding pocket of the kinase; contacting the cell with an ATP analog-nanoparticle conjugate capable of intracellular delivery of the ATP analog-nanoparticle conjugate, wherein the ATP analog comprises a detectable label; culturing the cells under conditions in which the ATP analog-nanoparticle conjugate contacts the one or more mutated kinases in cellulo, wherein the detectable label is transferred from the ATP analog-nanoparticle conjugate to a substrate of the one or more mutated kinases, wherein the one or more kinases react to transfer the detectable label to the substrate.

Disclosed is a method of measuring protein kinase activity, including a) attaching GMBS (N-[γ-maleimidobutyryloxy]sulfosuccinimide ester) to a base plate, b) attaching a substrate that reacts with a protein kinase to the base plate having GMBS attached thereto, thus manufacturing a kit for measuring protein kinase activity, c) introducing, to the kit, a mixture of a sample to be analyzed and a buffer including triton X-100, and d) probing phosphorylation of the substrate caused by the protein kinase contained in the sample, thereby measuring the activity of the protein kinase.

According to one embodiment, a vector set includes a first vector and a second vector. The first vector includes a transposase target sequence, a first promoter sequence ligated to downstream of the transposase target sequence, and a first reporter gene ligated to downstream of the first promoter sequence. The second vector includes a 5′-side transposase recognition sequence, a 3′-side transposase recognition sequence, and a first enhancer sequence arranged therebetween.