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A road map to evaluate the proteome-wide selectivity of covalent kinase inhibitors

2014; Nature Portfolio; Volume: 10; Issue: 9 Linguagem: Inglês

10.1038/nchembio.1582

1552-4469

Bryan Lanning, Landon R. Whitby, Melissa M. Dix, John Douhan, A. Gilbert, Erik C. Hett, Theodore Johnson, Chris Joslyn, John C. Kath, Sherry Niessen, Lee R. Roberts, Mark E. Schnute, Chu Wang, Jonathan J. Hulce, Baoxian Wei, Laurence O. Whiteley, Matthew M. Hayward, Benjamin F. Cravatt,

Monoclonal and Polyclonal Antibodies Research

ABPP combined with quantitative MS enabled identification of specific on- and off-targets of covalent kinase inhibitors. Modifications to inhibitors that alter specificity beyond a defined window can promote kinase-independent toxicity. 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, have 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 MS to globally map the targets, both specific and nonspecific, of covalent kinase inhibitors in human cells. Many of the specific off-targets represent nonkinase proteins that, notably, have 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 road map to illuminate opportunities and surmount challenges for the development of covalent kinase inhibitors.