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

<p>Protein <i>S</i>-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 <b>cyano-myracrylamide (CMA)</b>, 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 α-halo fatty acid, <b>CMA </b>inhibits DHHC family proteins <i>in cellulo</i> while demonstrating decreased toxicity and avoiding inhibition<b> </b>of the <i>S-</i>acylation eraser enzymes – two of the major weaknesses of 2BP. Our studies show that <b>CMA </b>engages with DHHC family proteins in cells, inhibits protein <i>S-</i>acylation, and disrupts DHHC-regulated cellular events. <b>CMA</b> represents an improved chemical scaffold for untangling the complexities of DHHC-mediated cell signaling by protein <i>S</i>-acylation.</p>