title: Dentipain, a Streptococcus pyogenes IdeS protease homolog, is a novel virulence factor of Treponema denticola.
abstract: Treponema denticola is a major pathogen of chronic periodontitis. Analysis of the T. denticola genome revealed a gene orthologous with a cysteine protease-encoding gene from Streptococcus pyogenes (IdeS). IdeS interferes with IgG-dependent opsonophagocytosis by specific cleavage of IgG molecules. Analysis of this gene (termed ideT) revealed it to encode a two-domain protein whose N-terminus is composed of tandem immunoglobulin-like domains followed by a C-terminal IdeS-like protease domain. In this study we show that during secretion the IdeT protein is processed into an N-terminal fragment which remains associated with the cell, and a C-terminal part released into the medium. Although the secreted domain of IdeT, termed dentipain, shows only 25% identity to the IdeS protease, the putative catalytic cysteine and histidine residues are strongly conserved. Recombinant dentipain cleaves the insulin β-chain, an activity which is inhibited by E-64, a diagnostic inhibitor of cysteine proteases. Apart from insulin no cleavage of other protein substrates was detected, suggesting that dentipain has oligopeptidase activity. A mutant strain was constructed expressing a modified IdeT variant, the dentipain domain of which was deleted. This strain was found to be significantly reduced in its abscess-forming activity compared with the parental strain in a murine abscess model, suggesting that dentipain contributes to the virulence of T. denticola.
authors.affiliation: Oral Health Science Center, Tokyo Dental College, Chiba 261-8502, Japan. ishihara@tdc.ac.jp
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Ishihara
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Wawrzonek
authors.forename: Lindesey N
authors.initials: LN
authors.lastname: Shaw
authors.forename: Satoru
authors.initials: S
authors.lastname: Inagaki
authors.forename: Meguru
authors.initials: M
authors.lastname: Miyamoto
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D048373:Treponema denticola / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D015853:Cysteine Proteinase Inhibitors; D037521:Virulence Factors; D012697:Serine Endopeptidases; D007930:Leucine; C024974:E 64