title: Profiling native kinases by immuno-assisted activity-based profiling.
abstract: The protocols in this unit describe efficient and cost-effective approaches to determine the interaction of small-molecule inhibitors with native kinases, and also analyze the interactions between kinases and their binding partners in a cellular setting. The combined attributes of activity-based probes and western blotting procedures provide for quantitative measurement of inhibitor efficacy, isoform selectivity, and post-translational modifications. We further demonstrate the ability to identify protein-protein interactions between a probe-labeled protein and its noncovalent binding partners.
authors.affiliation: ActivX Biosciences, La Jolla, California.
authors.forename: Eric
authors.initials: E
authors.lastname: Okerberg
authors.forename: Ann
authors.initials: A
authors.lastname: Shih
authors.forename: Heidi
authors.initials: H
authors.lastname: Brown
authors.forename: Senait
authors.initials: S
authors.lastname: Alemayehu
authors.forename: Arwin
authors.initials: A
authors.lastname: Aban
authors.forename: Tyzoon K
authors.initials: TK
authors.lastname: Nomanbhoy
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D066246:ErbB Receptors / Q000037:antagonists & inhibitors; D006801:Humans; D007158:Immunologic Techniques; D007202:Indicators and Reagents; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011494:Protein Kinases / Q000737:chemistry* / Q000276:immunology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D054852:Small Molecule Libraries
chemical_list: D007202:Indicators and Reagents; D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; D011494:Protein Kinases; D066246:ErbB Receptors; D048670:MAP Kinase Kinase 4
keywords: activity based protein profiling; immuno-assisted activity based profiling; inhibitors; native kinases