Broad-spectrum kinase profiling in live cells with lysine-targeted sulfonyl fluoride probes

Qian Zhao, Xiaohu Ouyang, Xiaobo Wan, Ketan S. Gajiwala, John C. Kath, Lyn H. Jones, Alma L. Burlingame, Jack Taunton

Research output: Journal article publicationJournal articleAcademic researchpeer-review

103 Citations (Scopus)

Abstract

Protein kinases comprise a large family of structurally related enzymes. A major goal in kinase-inhibitor development is to selectively engage the desired kinase while avoiding myriad off-target kinases. However, quantifying inhibitor interactions with multiple endogenous kinases in live cells remains an unmet challenge. Here, we report the design of sulfonyl fluoride probes that covalently label a broad swath of the intracellular kinome with high efficiency. Protein crystallography and mass spectrometry confirmed a chemo-selective reaction between the sulfonyl fluoride and a conserved lysine in the ATP binding site. Optimized probe 2 (XO44) covalently modified up to 133 endogenous kinases, efficiently competing with high intracellular concentrations of ATP. We employed probe 2 and label-free mass spectrometry to quantify intracellular kinase engagement by the approved drug, dasatinib. The data revealed saturable dasatinib binding to a small subset of kinase targets at clinically relevant concentrations, highlighting the utility of lysine-targeted sulfonyl fluoride probes in demanding chemoproteomic applications.
Original languageEnglish
Pages (from-to)680-685
Number of pages6
JournalJournal of the American Chemical Society
Volume139
Issue number2
DOIs
Publication statusPublished - 18 Jan 2017
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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