title: Clostridium difficile sortase recognizes a (S/P)PXTG sequence motif and can accommodate diaminopimelic acid as a substrate for transpeptidation.
abstract: Covalent attachment of surface proteins to the cell wall of Gram-positive bacteria requires a sortase-mediated transpeptidation reaction. In almost all Gram-positive bacteria, the housekeeping sortase, sortase A, recognizes the canonical recognition sequence LPXTG (X=any amino acid). The human pathogen Clostridium difficile carries a single putative sortase gene (cd2718) but neither transpeptidation activity nor specificity of CD2718 has been investigated. We produced recombinant CD2718 and examined its transpeptidation activity in vitro using synthetic peptides and MALDI-ToF(-ToF) MS analysis. We demonstrate that CD2718 has sortase activity with specificity for a (S/P)PXTG motif and can accommodate diaminopimelic acid as a substrate for transpeptidation.
authors.affiliation: Department of Medical Microbiology, Section Experimental Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Hans C
authors.initials: HC
authors.lastname: van Leeuwen
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Oleg I
authors.initials: OI
authors.lastname: Klychnikov
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Mica A C
authors.initials: MA
authors.lastname: Menks
authors.affiliation: Department of Medical Microbiology, Section Experimental Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Ed J
authors.initials: EJ
authors.lastname: Kuijper
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands.
authors.forename: Jan W
authors.initials: JW
authors.lastname: Drijfhout
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands. Electronic address: P.Hensbergen@lumc.nl.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hensbergen
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D016360:Clostridioides difficile / Q000201:enzymology*; D019295:Computational Biology; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D003960:Diaminopimelic Acid / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D011994:Recombinant Proteins; D003960:Diaminopimelic Acid; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: Adhesion; Diaminopimelic acid; Mass spectrometry; Peptidoglycan; Transpeptidation; Virulence