1. Academic Validation
  2. A road map to evaluate the proteome-wide selectivity of covalent kinase inhibitors

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

  • Nat Chem Biol. 2014 Sep;10(9):760-767. doi: 10.1038/nchembio.1582.
Bryan R Lanning  # 1 Landon R Whitby  # 1 Melissa M Dix 1 John Douhan 2 Adam M Gilbert 3 Erik C Hett 2 Theodore O Johnson 4 Chris Joslyn 1 John C Kath 4 Sherry Niessen 4 Lee R Roberts 2 Mark E Schnute 2 Chu Wang 1 Jonathan J Hulce 1 Baoxian Wei 5 Laurence O Whiteley 3 Matthew M Hayward 3 Benjamin F Cravatt 1
Affiliations

Affiliations

  • 1 The Skaggs Institute for Chemical Biology and Department of Chemical Physiology, The Scripps Research Institute, 10550 N. Torrey Pines Rd. La Jolla, CA, 92307.
  • 2 Pfizer Worldwide Research and Development, 200 Cambridge Park Drive, Cambridge, MA 02140.
  • 3 Pfizer Worldwide Research and Development, Eastern Point Road, Groton, CT 06340.
  • 4 Pfizer Worldwide Research and Development, 10770 Science Park Drive, San Diego, CA 92121.
  • 5 Pfizer Worldwide Research and Development, 1 Burtt Rd, Andover, MA 01810.
  • # Contributed equally.
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, 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.

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