1. Reversible lysine-targeted probes reveal residence time-based kinase selectivity
- Author
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Yang, Tangpo, Cuesta, Adolfo, Wan, Xiaobo, Craven, Gregory B, Hirakawa, Brad, Khamphavong, Penney, May, Jeffrey R, Kath, John C, Lapek, John D, Niessen, Sherry, Burlingame, Alma L, Carelli, Jordan D, and Taunton, Jack
- Subjects
Biochemistry and Cell Biology ,Chemical Sciences ,Biological Sciences ,Brain Disorders ,Animals ,Cysteine ,Lysine ,Mice ,Protein Binding ,Protein Kinase Inhibitors ,Protein Kinases ,Medicinal and Biomolecular Chemistry ,Biochemistry & Molecular Biology ,Biochemistry and cell biology ,Medicinal and biomolecular chemistry - Abstract
The expansion of the target landscape of covalent inhibitors requires the engagement of nucleophiles beyond cysteine. Although the conserved catalytic lysine in protein kinases is an attractive candidate for a covalent approach, selectivity remains an obvious challenge. Moreover, few covalent inhibitors have been shown to engage the kinase catalytic lysine in animals. We hypothesized that reversible, lysine-targeted inhibitors could provide sustained kinase engagement in vivo, with selectivity driven in part by differences in residence time. By strategically linking benzaldehydes to a promiscuous kinase binding scaffold, we developed chemoproteomic probes that reversibly and covalently engage >200 protein kinases in cells and mice. Probe-kinase residence time was dramatically enhanced by a hydroxyl group ortho to the aldehyde. Remarkably, only a few kinases, including Aurora A, showed sustained, quasi-irreversible occupancy in vivo, the structural basis for which was revealed by X-ray crystallography. We anticipate broad application of salicylaldehyde-based probes to proteins that lack a druggable cysteine.
- Published
- 2022