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

ACS Chem Biol. 2021 Aug 20;16(8):1546-1556. doi: 10.1021/acschembio.1c00405. Epub 2021 Jul 26.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemical synthesis
  • Acrylamides / pharmacology*
  • Acrylamides / toxicity
  • Acylation / drug effects
  • Acyltransferases / antagonists & inhibitors*
  • Animals
  • CD36 Antigens / metabolism*
  • Cell Line, Tumor
  • Enzyme Inhibitors / chemical synthesis
  • Enzyme Inhibitors / pharmacology*
  • Enzyme Inhibitors / toxicity
  • ErbB Receptors / metabolism
  • Humans
  • Lipoylation / drug effects
  • Mice
  • Protein Processing, Post-Translational / drug effects

Substances

  • Acrylamides
  • CD36 Antigens
  • Cd36 protein, mouse
  • Enzyme Inhibitors
  • Acyltransferases
  • ZDHHC20 protein, human
  • EGFR protein, human
  • ErbB Receptors