Structures of human peptidylarginine deiminase type III provide insights into substrate recognition and inhibitor design

• Published in: Archives of biochemistry and biophysics 2021-09, Vol.708, p.108911, Article 108911
• Main Authors: Funabashi, Kazumasa, Sawata, Mizuki, Nagai, Anna, Akimoto, Megumi, Mashimo, Ryutaro, Takahara, Hidenari, Kizawa, Kenji, Thompson, Paul R., Ite, Kenji, Kitanishi, Kenichi, Unno, Masaki
• Format: Article
• Language: English
• Subjects: Amidines - chemistry; Amidines - pharmacology; Amino Acid Sequence; Catalytic Domain; Citrulline; Crystallography, X-Ray; Drug Design; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Filaggrin Proteins; Humans; Hydrolases - antagonists & inhibitors; Hydrolases - chemistry; Hydrolases - metabolism; Inhibitor; Isozyme; Models, Molecular; Ornithine - analogs & derivatives; Post-translational modification; Protein Conformation; Protein-Arginine Deiminase Type 3 - chemistry; Protein-Arginine Deiminase Type 3 - metabolism; Protein-Arginine Deiminase Type 4 - chemistry; Protein-Arginine Deiminase Type 4 - metabolism; Protein-Arginine Deiminases - antagonists & inhibitors; Protein-Arginine Deiminases - chemistry; Protein-Arginine Deiminases - metabolism; Quaternary structure; S100 Proteins - chemistry; S100 Proteins - metabolism; Selectivity; Substrate Specificity
• ISSN: 0003-9861; 1096-0384; 1096-0384
• DOI: 10.1016/j.abb.2021.108911

Peptidylarginine deiminase type III (PAD3) is an isozyme belonging to the PAD enzyme family that converts arginine to citrulline residue(s) within proteins. PAD3 is expressed in most differentiated keratinocytes of the epidermis and hair follicles, while S100A3, trichohyalin, and filaggrin are its principal substrates. In this study, the X-ray crystal structures of PAD3 in six states, including its complex with the PAD inhibitor Cl-amidine, were determined. This structural analysis identified a large space around Gly374 in the PAD3-Ca2+-Cl-amidine complex, which may be used to develop novel PAD3-selective inhibitors. In addition, similarities between PAD3 and PAD4 were found based on the investigation of PAD4 reactivity with S100A3 in vitro. A comparison of the structures of PAD1, PAD2, PAD3, and PAD4 implied that the flexibility of the structures around the active site may lead to different substrate selectivity among these PAD isozymes.