Development of an Acrylamide-Based Inhibitor of Protein S-Acylation

Protein S-acylation is a dynamic lipid post-translational modification that can modulate the localization and activity of target proteins. In humans, the installation of the lipid onto target proteins is catalyzed by a family of 23 Asp-His-His-Cys domain-containing protein acyltransferases (DHHC-PATs). DHHCs are increasingly recognized as critical players in cellular signaling events and in human disease. However, progress elucidating the functions and mechanisms of DHHC writers has been hampered by a lack of chemical tools to perturb their activity in live cells. Herein, we report the synthesis and characterization of cyano-myracrylamide (CMA), a broad-spectrum DHHC family inhibitor with similar potency to 2-bromopalmitate (2BP), the most commonly used DHHC inhibitor in the field. Possessing an acrylamide warhead instead of 2BP's alpha-halo fatty acid, CMA inhibits DHHC family proteins in cellulo while demonstrating decreased toxicity and avoiding inhibition of the S-acylation eraser enzymes, two of the major weaknesses of 2BP. Our studies show that CMA engages with DHHC family proteins in cells, inhibits protein S-acylation, and disrupts DHHC-regulated cellular events. CMA represents an improved chemical scaffold for untangling the complexities of DHHC-mediated cell signaling by protein S-acylation.

Automated summary

Protein S-acylation is a dynamic lipid post-translational modification that can modulate the localization and activity of target proteins. A new broad-spectrum DHHC family inhibitor called cyano-myracrylamide (CMA) was synthesized and characterized, showing potent inhibition of DHHC family proteins with decreased toxicity and avoidance of inhibiting the S-acylation eraser enzymes. CMA provides an improved chemical scaffold for understanding the complexities of DHHC-mediated cell signaling by protein S-acylation.