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.

中文翻译：

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基于丙烯酰胺的蛋白质 S-酰化抑制剂的开发

蛋白质S-酰化是一种动态脂质翻译后修饰，可以调节靶蛋白的定位和活性。在人类中，脂质在靶蛋白上的安装由 23 个含有 Asp-His-His-Cys 结构域的蛋白酰基转移酶 (DHHC-PAT) 家族催化。DHHC 越来越被认为是细胞信号传导事件和人类疾病中的关键参与者。然而，由于缺乏化学工具来干扰其在活细胞中的活动，阐明 DHHC“作家”的功能和机制的进展受到了阻碍。在此，我们报道了氰基-丙烯酰胺（ CMA）的合成和表征。)，一种广谱 DHHC 家族抑制剂，与该领域最常用的 DHHC 抑制剂 2-溴棕榈酸酯 (2BP) 具有相似的效力。CMA拥有丙烯酰胺弹头而不是 2BP 的 α-卤代脂肪酸，可抑制纤维素中的 DHHC 家族蛋白，同时降低毒性并避免抑制S-酰化擦除酶，这是 2BP 的两个主要弱点。我们的研究表明，CMA与细胞中的 DHHC 家族蛋白结合，抑制蛋白S-酰化，并破坏 DHHC 调节的细胞事件。CMA代表了一种改进的化学支架，用于解开由蛋白质S引起的 DHHC 介导的细胞信号传导的复杂性-酰化。