Fluorescence linked enzyme chemoproteomic strategy for discovery of a potent and selective DAPK1 and ZIPK inhibitor.

Journal Article (Journal Article)

DAPK1 and ZIPK (also called DAPK3) are closely related serine/threonine protein kinases that regulate programmed cell death and phosphorylation of non-muscle and smooth muscle myosin. We have developed a fluorescence linked enzyme chemoproteomic strategy (FLECS) for the rapid identification of inhibitors for any element of the purinome and identified a selective pyrazolo[3,4-d]pyrimidinone (HS38) that inhibits DAPK1 and ZIPK in an ATP-competitive manner at nanomolar concentrations. In cellular studies, HS38 decreased RLC20 phosphorylation. In ex vivo studies, HS38 decreased contractile force generated in mouse aorta, rabbit ileum, and calyculin A stimulated arterial muscle by decreasing RLC20 and MYPT1 phosphorylation. The inhibitor also promoted relaxation in Ca(2+)-sensitized vessels. A close structural analogue (HS43) with 5-fold lower affinity for ZIPK produced no effect on cells or tissues. These findings are consistent with a mechanism of action wherein HS38 specifically targets ZIPK in smooth muscle. The discovery of HS38 provides a lead scaffold for the development of therapeutic agents for smooth muscle related disorders and a chemical means to probe the function of DAPK1 and ZIPK across species.

Full Text

Duke Authors

Cited Authors

  • Carlson, DA; Franke, AS; Weitzel, DH; Speer, BL; Hughes, PF; Hagerty, L; Fortner, CN; Veal, JM; Barta, TE; Zieba, BJ; Somlyo, AV; Sutherland, C; Deng, JT; Walsh, MP; MacDonald, JA; Haystead, TAJ

Published Date

  • December 20, 2013

Published In

Volume / Issue

  • 8 / 12

Start / End Page

  • 2715 - 2723

PubMed ID

  • 24070067

Pubmed Central ID

  • PMC4445880

Electronic International Standard Serial Number (EISSN)

  • 1554-8937

Digital Object Identifier (DOI)

  • 10.1021/cb400407c


  • eng

Conference Location

  • United States