1. A roadmap to evaluate the proteome-wide selectivity of covalent kinase inhibitors
- Author
-
John Charles Kath, Theodore O. Johnson, Landon R. Whitby, Benjamin F. Cravatt, Baoxian Wei, Chu Wang, Mark E. Schnute, Melissa M. Dix, Lee R. Roberts, Laurence O. Whiteley, Adam M. Gilbert, Jonathan J. Hulce, Sherry Niessen, Erik C. Hett, Chris Joslyn, Matthew Merrill Hayward, Bryan R. Lanning, and John Douhan
- Subjects
Proteome ,Cell Survival ,Proteomics ,01 natural sciences ,Article ,03 medical and health sciences ,Piperidines ,Cell Line, Tumor ,Agammaglobulinaemia Tyrosine Kinase ,Humans ,Cysteine ,Molecular Biology ,Protein Kinase Inhibitors ,030304 developmental biology ,0303 health sciences ,biology ,010405 organic chemistry ,Kinase ,Drug discovery ,Adenine ,Active site ,Cell Biology ,Genes, erbB-1 ,Protein-Tyrosine Kinases ,Small molecule ,0104 chemical sciences ,Kinetics ,Pyrimidines ,Biochemistry ,Covalent bond ,biology.protein ,Pyrazoles ,Signal transduction ,Protein Kinases ,Signal Transduction - Abstract
Kinases are principal components of signal transduction pathways and the focus of intense basic and drug discovery research. Irreversible inhibitors that covalently modify non-catalytic cysteines in kinase active-sites have emerged as valuable probes and approved drugs. Many protein classes, however, possess functional cysteines and therefore understanding the proteome-wide selectivity of covalent kinase inhibitors is imperative. Here, we accomplish this objective using activity-based protein profiling coupled with quantitative mass spectrometry to globally map the targets, both specific and non-specific, of covalent kinase inhibitors in human cells. Many of the specific off-targets represent non-kinase proteins that, interestingly, possess conserved, active-site cysteines. We define windows of selectivity for covalent kinase inhibitors and show that, when these windows are exceeded, rampant proteome-wide reactivity and kinase target-independent cell death conjointly occur. Our findings, taken together, provide an experimental roadmap to illuminate opportunities and surmount challenges for the development of covalent kinase inhibitors.
- Published
- 2014