title: Kinase inhibitor profiling using chemoproteomics.
abstract: 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.
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA, USA. markus.schirle@novartis.com
authors.forename: Markus
authors.initials: M
authors.lastname: Schirle
authors.forename: Eugene C
authors.initials: EC
authors.lastname: Petrella
authors.forename: Scott M
authors.initials: SM
authors.lastname: Brittain
authors.forename: David
authors.initials: D
authors.lastname: Schwalb
authors.forename: Edmund
authors.initials: E
authors.lastname: Harrington
authors.forename: Ivan
authors.initials: I
authors.lastname: Cornella-Taracido
authors.forename: John A
authors.initials: JA
authors.lastname: Tallarico
mesh_terms: D001667:Binding, Competitive; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D013058:Mass Spectrometry; D010455:Peptides / Q000378:metabolism; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D004791:Enzyme Inhibitors; D010455:Peptides; D010770:Phosphotransferases