title: Clostridium difficile secreted Pro-Pro endopeptidase PPEP-1 (ZMP1/CD2830) modulates adhesion through cleavage of the collagen binding protein CD2831.
abstract: The Clostridium difficile cd2830 gene product is a secreted metalloprotease, named Pro-Pro endopeptidase (PPEP-1). PPEP-1 cleaves C. difficile cell surface proteins (e.g. CD2831). Here, we confirmed that PPEP-1 has a unique preference for prolines surrounding the scissile bond. Moreover, we show that it exhibits a high preference for an asparagine at the P2 position and hydrophobic residues at the P3 position. Using a PPEP-1 knockout C. difficile strain, we demonstrate that the removal of the collagen binding protein CD2831 is fully attributable to PPEP-1 activity. The PPEP-1 knockout strain demonstrated higher affinity for collagen type I with attenuated virulence in hamsters.
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands. Electronic address: P.Hensbergen@lumc.nl.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hensbergen
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands.
authors.forename: Oleg I
authors.initials: OI
authors.lastname: Klychnikov
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands.
authors.forename: Dennis
authors.initials: D
authors.lastname: Bakker
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands.
authors.forename: Irina
authors.initials: I
authors.lastname: Dragan
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, The University of Nottingham, NG7 2RD, United Kingdom.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Kelly
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, The University of Nottingham, NG7 2RD, United Kingdom.
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Minton
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Corver
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, PO Box 9600, 2300RC 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 Wouter
authors.initials: JW
authors.lastname: Drijfhout
authors.affiliation: Department of Medical Microbiology, Section Experimental Bacteriology, Leiden University Medical Center, PO Box 9600, 2300RC Leiden, The Netherlands. Electronic address: H.C.van_Leeuwen@lumc.nl.
authors.forename: Hans C
authors.initials: HC
authors.lastname: van Leeuwen
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D016360:Clostridioides difficile / Q000201:enzymology / Q000472:pathogenicity / Q000502:physiology*; D024042:Collagen Type I / Q000378:metabolism*; D005260:Female; D055786:Gene Knockout Techniques; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008647:Mesocricetus; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D059748:Proteolysis; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D024042:Collagen Type I; D008565:Membrane Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D008666:Metalloendopeptidases
keywords: Adhesion; Bacterial virulence; Collagen binding; Motility; Secreted protease