on 05-01-201210:50 AM - edited on 10-15-202111:41 AM by Closed Account
Schirle M, Petrella EC, Brittain SM, Schwalb D, Harrington E, Cornella-Taracido I, Tallarico JA. Methods Mol Biol. 2012;795:161-77. Quantitative chemoproteomics has recently emerged as an experimental approach to determine protein interaction profiles of small molecules in a given cell line or tissue. In contrast to standard biochemical and biophysical kinase assays, application of this method to kinase inhibitors determines compound binding to endogenously expressed kinases under conditions approximating the physiological situation with regard to the molecular state of the kinase and presence of required cofactors and regulatory proteins. Using a dose-dependent, competition-based experimental design in combination with quantitative mass spectrometry approaches, such as the use of tandem mass tags (TMT) for isobaric labeling described here, allows to rank-order interactions of inhibitors to kinase by binding affinity.