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.
Keywords:
Adhesion; Bacterial virulence; Collagen binding; Motility; Secreted protease.
Copyright © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
MeSH terms
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Amino Acid Motifs
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Animals
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Bacterial Adhesion*
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Bacterial Proteins / chemistry
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Bacterial Proteins / metabolism*
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Clostridioides difficile / enzymology
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Clostridioides difficile / pathogenicity
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Clostridioides difficile / physiology*
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Collagen Type I / metabolism*
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Female
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Gene Knockout Techniques
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Hydrophobic and Hydrophilic Interactions
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Mesocricetus
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Metalloendopeptidases / genetics
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Metalloendopeptidases / metabolism*
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Mutation
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Protein Interaction Domains and Motifs
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Proteolysis
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Rats
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Substrate Specificity
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Virulence
Substances
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Bacterial Proteins
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Collagen Type I
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Membrane Proteins
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Peptide Fragments
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Recombinant Fusion Proteins
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Metalloendopeptidases