The RgpB C-Terminal Domain Has a Role in Attachment of RgpB to the Outer Membrane and Belongs to a Novel C-Terminal-Domain Family Found in Porphyromonas gingivalis
Christine A. Seers,Nada Slakeski,Paul D. Veith,Todd Nikolof,Yu-Yen Chen,Stuart G. Dashper,Eric C. Reynolds +6 more
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TLDR
The results suggest that the C-terminal domain is essential for outer membrane attachment and may be involved in a coordinated process of export and attachment to the cell surface.Abstract:
Porphyromonas gingivalis produces outer membrane-attached proteins that include the virulence-associated proteinases RgpA and RgpB (Arg-gingipains) and Kgp (Lys-gingipain). We analyzed the P. gingivalis outer membrane proteome and identified numerous proteins with C-terminal domains similar in sequence to those of RgpB, RgpA, and Kgp, indicating that these domains may have a common function. Using RgpB as a model to investigate the role of the C-terminal domain, we expressed RgpB as a full-length zymogen (recombinant RgpB [rRgpB]), with a catalytic Cys244Ala mutation [rRgpB(C244A)], or with the C-terminal 72 amino acids deleted (rRgpB435) in an Arg-gingipain P. gingivalis mutant (YH522AB) and an Arg- and Lys-gingipain mutant (YH522KAB). rRgpB was catalytically active and located predominantly attached to the outer membrane of both background strains. rRgpB(C244A) was inactive and outer membrane attached, with a typical attachment profile for both background strains according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but in YH522KAB, the prodomain was not removed. Thus, in vivo, RgpB export and membrane attachment are independent of the proteolytic activity of RgpA, RgpB, or Kgp. However, for maturation involving proteolytic processing of RgpB, the proteolytic activity of RgpB, RgpA, or Kgp is required. The C-terminally-truncated rRgpB435 was not attached to the outer membrane and was located as largely inactive, discrete 71-kDa and 48-kDa isoforms in the culture supernatant and the periplasm. These results suggest that the C-terminal domain is essential for outer membrane attachment and may be involved in a coordinated process of export and attachment to the cell surface.read more
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Genomic characterization of the uncultured Bacteroidales family S24-7 inhabiting the guts of homeothermic animals
Kate L. Ormerod,David L. A. Wood,Nancy Lachner,Shaan L. Gellatly,Joshua Daly,Jeremy Parsons,Cristiana Gomes de Oliveira Dal'Molin,Robin W. Palfreyman,Lars K. Nielsen,Mark E. Cooper,Mark Morrison,Philip M. Hansbro,Philip Hugenholtz +12 more
TL;DR: The first whole genome exploration of the Bacteroidales family S24-7 is provided, for which the name “Candidatus Homeothermaceae” is proposed, using 30 population genomes extracted from fecal samples of four different animal hosts.
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A protein secretion system linked to bacteroidete gliding motility and pathogenesis
Keiko Sato,Mariko Naito,Hideharu Yukitake,Hideki Hirakawa,Mikio Shoji,Mark J. McBride,Ryan G. Rhodes,Koji Nakayama +7 more
TL;DR: Real-time RT-PCR analysis revealed that porK, porL, porM, porN, porP, porT, and sov were down-regulated in P. gingivalis porX and porY mutants, providing a link between a unique protein translocation system and a motility apparatus in members of the Bacteroidetes phylum.
Journal ArticleDOI
Dichotomy of gingipains action as virulence factors: from cleaving substrates with the precision of a surgeon’s knife to a meat chopper‐like brutal degradation of proteins
TL;DR: Gingipains are absolutely essential for bacterial survival/proliferation in vivo and for the pathological outcome of the experimental infection of P. gingivalis, the most prevailing form of periodontal disease.
Journal ArticleDOI
Gliding motility and Por secretion system genes are widespread among members of the phylum bacteroidetes.
Mark J. McBride,Yongtao Zhu +1 more
TL;DR: Comparative analysis of 37 genomes of members of the phylum Bacteroidetes revealed the widespread occurrence of gliding motility genes and PorSS genes, suggesting that glider motility is more common than previously reported.
Journal ArticleDOI
Porphyromonas gingivalis outer membrane vesicles exclusively contain outer membrane and periplasmic proteins and carry a cargo enriched with virulence factors.
Paul D. Veith,Yu-Yen Chen,Dhana G. Gorasia,Dina Chen,Michelle D. Glew,Neil M O'Brien-Simpson,Jessica D. Cecil,James A Holden,Eric C. Reynolds +8 more
TL;DR: Cryo-transmission electron microscopy analysis revealed that an electron dense surface layer known to comprise CTD proteins accounted for a large proportion of the OMVs' volume providing an explanation for the enrichment of CTDprotein, finding that P. gingivalis is able to specifically concentrate and release a large number of its virulence factors into the environment in the form of OMVs.
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