5GYJ

Structure of catalytically active sortase from Clostridium difficile

Summary for 5GYJ

• Entry DOI: 10.2210/pdb5gyj/pdb
• Related: 4UX7
• Descriptor: Putative peptidase C60B, sortase B (2 entities in total)
• Functional Keywords: transpeptidase, cys-his-arg catalytic triad, hydrolase
• Biological source: Peptoclostridium difficile (strain 630)
• Total number of polymer chains: 1
• Total formula weight: 26189.28

Authors

Yin, J.-C.,Fei, C.-H.,Hsiao, Y.-Y.,Nix, J.C.,Huang, I.-H.,Wang, S. (deposition date: 2016-09-22, release date: 2017-01-04, Last modification date: 2023-11-08)

Primary citation

Yin, J.C.,Fei, C.H.,Lo, Y.C.,Hsiao, Y.Y.,Chang, J.C.,Nix, J.C.,Chang, Y.Y.,Yang, L.W.,Huang, I.H.,Wang, S.
Structural Insights into Substrate Recognition by Clostridium difficile Sortase.
Front Cell Infect Microbiol, 6:160-160, 2016

PubMed Abstract: 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 sortase B (Cd-SrtB). Combining structural studies, biochemical assays and molecular dynamics simulations, we have constructed a computational model of Cd-SrtB-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.

PubMed: 27921010
DOI: 10.3389/fcimb.2016.00160

Experimental method

X-RAY DIFFRACTION (2.801 Å)