Structural insights into substrate recognition by Clostridium difficile sortase

Jui Chieh Yin, Chun Hsien Fei, Yen Chen Lo, Yu Yuan Hsiao, Jyun Cyuan Chang, Jay C. Nix, Yuan Yu Chang, Lee Wei Yang, I. Hsiu Huang, Shuying Wang

研究成果: Article同行評審

4 引文 斯高帕斯(Scopus)

摘要

Sortases function as cysteine transpeptidases that catalyze the covalent attachment of virulence-associated surface proteins into the cell wall peptidoglycan in Gram-positive bacteria. The substrate proteins targeted by sortase enzymes have a cell wall sorting signal (CWSS) located at the C-terminus. Up to date, it is still not well understood how sortases with structural resemblance among different classes and diverse species of bacteria achieve substrate specificity. In this study, we focus on elucidating the molecular basis for specific recognition of peptide substrate PPKTG by Clostridium difficile sortase B (Cd-SrtB). Combining structural studies, biochemical assays and molecular dynamics simulations, we have constructed a computational model of Cd-SrtBΔN26-PPKTG complex and have validated the model by site-directed mutagensis studies and fluorescence resonance energy transfer (FRET)-based assay. Furthermore, we have revealed that the fourth amino acid in the N-terminal direction from cleavage site of PPKTG forms specific interaction with Cd-SrtB and plays an essential role in configuring the peptide to allow more efficient substrate-specific cleavage by Cd-SrtB.

原文English
文章編號160
期刊Frontiers in Cellular and Infection Microbiology
6
發行號NOV
DOIs
出版狀態Published - 2016 11月 22

All Science Journal Classification (ASJC) codes

  • 微生物學
  • 免疫學
  • 微生物學(醫學)
  • 傳染性疾病

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