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Crystal Structure of Streptococcus pyogenes Sortase A: Implications for Sortase mechanism

DOI: 10.1074/jbc.M805406200 DOI Help
PMID: 19129180 PMID Help

Authors: Paul Race (Newcastle University) , M L Bentley (Duke University, Durham, North Carolina) , Jeff Melvin (Duke University, Durham, North Carolina) , Allister Crow (Department of Biological Chemistry, John Innes Centre, Colney Lane, Norwich) , Richard Hughes (John Innes Centre) , Wendy Smith (University of Newcastle) , Richard Sessions (Department of Biochemistry, School of Medical Sciences, University of Bristol) , Michael Kehoe (Newcastle University) , Dewey Mccafferty (Duke University, Durham, North Carolina) , Mark Banfield (Newcastle University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Biological Chemistry , VOL 284 (11) , PAGES 6924-6933

State: Published (Approved)
Published: January 2009

Abstract: Sortases are a family of Gram-positive bacterial transpeptidases that anchor secreted proteins to bacterial cell surfaces. These include many proteins that play critical roles in the virulence of Gram-positive bacterial pathogens such that sortases are attractive targets for development of novel antimicrobial agents. All Gram-positive pathogens express a “housekeeping” sortase that recognizes the majority of secreted proteins containing an LPXTG wall-sorting motif and covalently attaches these to bacterial cell wall peptidoglycan. Many Gram-positive pathogens also express additional sortases that link a small number of proteins, often with variant wall-sorting motifs, to either other surface proteins or peptidoglycan. To better understand the mechanisms of catalysis and substrate recognition by the housekeeping sortase produced by the important human pathogen Streptococcus pyogenes, the crystal structure of this protein has been solved and its transpeptidase activity established in vitro. The structure reveals a novel arrangement of key catalytic residues in the active site of a sortase, the first that is consistent with kinetic analysis. The structure also provides a complete description of residue positions surrounding the active site, overcoming the limitation of localized disorder in previous structures of sortase A-type proteins. Modification of the active site Cys through oxidation to its sulfenic acid form or by an alkylating reagent supports a role for a reactive thiol/thiolate in the catalytic mechanism. These new insights into sortase structure and function could have important consequences for inhibitor design.

Journal Keywords: Bacterial; Catalytic; Crystallography; X-Ray; Cysteine; Kinetics; Oxidation-Reduction; Protein; Tertiary; Streptococcus; Sulfenic Acids

Subject Areas: Biology and Bio-materials

Instruments: I03-Macromolecular Crystallography

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