A Photoaffinity Labeling-Based Chemoproteomics Strategy for Unbiased Target Deconvolution of Small Molecule Drug Candidates

The combination of photoaffinity labeling (PAL) and quantitative chemoproteomics enables the comprehensive, unbiased determination of protein interaction profiles to support target identification of bioactive small molecules. This approach is amenable to cells in culture and compatible with pharmaco...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1647; p. 1
Main Authors Thomas, Jason R, Brittain, Scott M, Lipps, Jennifer, Llamas, Luis, Jain, Rishi K, Schirle, Markus
Format Journal Article
LanguageEnglish
Published United States 01.01.2017
Subjects
Click Chemistry
Conductometry
Drug Design
GTP-Binding Proteins - analysis
HEK293 Cells
Humans
Mass Spectrometry
Photoaffinity Labels - chemistry
Proteomics - methods
Receptors, G-Protein-Coupled - analysis
Small Molecule Libraries - analysis
Chemoproteomics
Isobaric mass tags
Photoaffinity labeling
Quantitative proteomics
G-protein coupled receptor
Target identification
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Summary:The combination of photoaffinity labeling (PAL) and quantitative chemoproteomics enables the comprehensive, unbiased determination of protein interaction profiles to support target identification of bioactive small molecules. This approach is amenable to cells in culture and compatible with pharmacologically relevant transmembrane target classes like G-protein coupled receptors and ions channels which have been notoriously hard to access by conventional chemoproteomics approaches. Here, we describe a strategy that combines PAL probe titration and competition with excess parental compounds with the goal of enabling the identification of specific interactors as well as assessing the functional relevance of a binding event for the phenotype under investigation.
ISSN:1940-6029
DOI:10.1007/978-1-4939-7201-2_1