Discovery, Characterization, and Structure of a Cell Active PAD2 Inhibitor Acting through a Novel Allosteric Mechanism

ACS Chem Biol. 2024 Oct 18;19(10):2186-2197. doi: 10.1021/acschembio.4c00397. Epub 2024 Sep 24.

Abstract

Peptidyl arginine deiminases (PADs) are important enzymes in many diseases, especially those involving inflammation and autoimmunity. Despite many years of effort, developing isoform-specific inhibitors has been a challenge. We describe herein the discovery of a potent, noncovalent PAD2 inhibitor, with selectivity over PAD3 and PAD4, from a DNA-encoded library. The biochemical and biophysical characterization of this inhibitor and two noninhibitory binders indicated a novel, Ca2+ competitive mechanism of inhibition. This was confirmed via X-ray crystallographic analysis. Finally, we demonstrate that this inhibitor selectively inhibits PAD2 in a cellular context.

MeSH terms

  • Allosteric Regulation / drug effects
  • Calcium / metabolism
  • Crystallography, X-Ray
  • Drug Discovery
  • Enzyme Inhibitors* / chemistry
  • Enzyme Inhibitors* / pharmacology
  • Humans
  • Protein-Arginine Deiminase Type 2* / antagonists & inhibitors
  • Protein-Arginine Deiminase Type 2* / metabolism

Substances

  • Protein-Arginine Deiminase Type 2
  • Enzyme Inhibitors
  • PADI2 protein, human
  • Calcium