6KYC

Structure of the S207A mutant of Clostridium difficile sortase B

6KYC の概要

• エントリーDOI: 10.2210/pdb6kyc/pdb
• 関連するPDBエントリー: 5gyj
• 分子名称: Putative peptidase C60B, sortase B (2 entities in total)
• 機能のキーワード: sortase b, cysteine transpeptidase, clostridium difficile, hydrolase
• 由来する生物種: Peptoclostridium difficile 630
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 26173.28

構造登録者

Kang, C.Y.,Huang, I.H.,Wu, T.Y.,Chang, J.C.,Hsiao, Y.Y.,Cheng, C.H.,Tsai, W.J.,Hsu, K.C.,Wang, S.Y. (登録日: 2019-09-17, 公開日: 2020-02-19, 最終更新日: 2023-11-22)

主引用文献

Kang, C.Y.,Huang, I.H.,Chou, C.C.,Wu, T.Y.,Chang, J.C.,Hsiao, Y.Y.,Cheng, C.H.,Tsai, W.J.,Hsu, K.C.,Wang, S.
Functional analysis ofClostridium difficilesortase B reveals key residues for catalytic activity and substrate specificity.
J.Biol.Chem., 295:3734-3745, 2020

PubMed Abstract: Most of Gram-positive bacteria anchor surface proteins to the peptidoglycan cell wall by sortase, a cysteine transpeptidase that targets proteins displaying a cell wall sorting signal. Unlike other bacteria, , the major human pathogen responsible for antibiotic-associated diarrhea, has only a single functional sortase (SrtB). Sortase's vital importance in bacterial virulence has been long recognized, and sortase B (Cd-SrtB) has become an attractive therapeutic target for managing infection. A better understanding of the molecular activity of Cd-SrtB may help spur the development of effective agents against infection. In this study, using site-directed mutagenesis, biochemical and biophysical tools, LC-MS/MS, and crystallographic analyses, we identified key residues essential for Cd-SrtB catalysis and substrate recognition. To the best of our knowledge, we report the first evidence that a conserved serine residue near the active site participates in the catalytic activity of Cd-SrtB and also SrtB from The serine residue indispensable for SrtB activity may be involved in stabilizing a thioacyl-enzyme intermediate because it is neither a nucleophilic residue nor a substrate-interacting residue, based on the LC-MS/MS data and available structural models of SrtB-substrate complexes. Furthermore, we also demonstrated that residues 163-168 located on the β6/β7 loop of Cd-SrtB dominate specific recognition of the peptide substrate PPKTG. The results of this work reveal key residues with roles in catalysis and substrate specificity of Cd-SrtB.

PubMed: 32005667
DOI: 10.1074/jbc.RA119.011322

実験手法

X-RAY DIFFRACTION (2.604 Å)