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

• Published in: ACS chemical biology Vol. 16; no. 8; pp. 1546 - 1556
• Main Authors: Azizi, Saara-Anne, Lan, Tong, Delalande, Clémence, Kathayat, Rahul S, Banales Mejia, Fernando, Qin, Alice, Brookes, Noah, Sandoval, Perla Jasmine, Dickinson, Bryan C
• Format: Journal Article
• Language: English
• Published: American Chemical Society 20-08-2021
• ISSN: 1554-8929; 1554-8937
• DOI: 10.1021/acschembio.1c00405

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 α-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.