Identification of a new membrane-associated protein that influences transport/maturation of gingipains and adhesins of Porphyromonas gingivalis

Keiko Sato, Eiko Sakai, Paul D. Veith, Mikio Shoji, Yuichiro Kikuchi, Hideharu Yukitake, Naoya Oohara, Mariko Naito, Kuniaki Okamoto, Eric C. Reynolds, Koji Nakayama

Research output: Contribution to journalArticle

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Abstract

The dual membrane envelopes of Gram-negative bacteria provide two barriers of unlike nature that regulate the transport of molecules into and out of organisms. Organisms have developed several systems for transport across the inner and outer membranes. The Gram-negative periodontopathogenic bacterium Porphyromonas gingivalis produces proteinase and adhesin complexes, gingipains/adhesins, on the cell surface and in the extracellular milieu as one of the major virulence factors. Gingipains and/or adhesins are encoded by kgp, rgpA, rgpB, and hagA on the chromosome. In this study, we isolated a P. gingivalis mutant (porT), which showed very weals activities of gingipains im the cell lysates and culture supernatants. Subcellular fractionation and immunoblot analysis demonstrated that precursor forms of gingipains and adhesins were accumulated in the periplasmic space of the porT mutant cells. Peptide mass fingerprinting and N-terminal amino acid sequencing of the precursor proteins and the kgp'-'rgpE chimera gene product in the porT mutant indicated that these proteins lacked the signal peptide regions, consistent with their accumulation in the periplasm. The PorT protein seemed to be membrane-associated and exposed to the periplasmic space, as revealed by subcellular fractionation and immunoblot analysis using anti-PorT antiserum. These results suggest that the membrane-associated protein PorT is essential for transport of the kgp, rgpA, rgpB, and hagA gene products across the outer membrane from the periplasm to the cell surface, where they are processed and matured.

Original languageEnglish
Pages (from-to)8668-8677
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number10
DOIs
Publication statusPublished - Mar 11 2005
Externally publishedYes

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Periplasm
Porphyromonas gingivalis
Protein Transport
Membrane Proteins
Membranes
Fractionation
Gram-Negative Bacteria
Bacteria
Genes
Peptide Mapping
Protein Precursors
Protein Sequence Analysis
Virulence Factors
Mutant Proteins
Chromosomes
Protein Sorting Signals
Immune Sera
Proteins
Peptide Hydrolases
Cell Culture Techniques

ASJC Scopus subject areas

  • Biochemistry

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Identification of a new membrane-associated protein that influences transport/maturation of gingipains and adhesins of Porphyromonas gingivalis. / Sato, Keiko; Sakai, Eiko; Veith, Paul D.; Shoji, Mikio; Kikuchi, Yuichiro; Yukitake, Hideharu; Oohara, Naoya; Naito, Mariko; Okamoto, Kuniaki; Reynolds, Eric C.; Nakayama, Koji.

In: Journal of Biological Chemistry, Vol. 280, No. 10, 11.03.2005, p. 8668-8677.

Research output: Contribution to journalArticle

Sato, Keiko ; Sakai, Eiko ; Veith, Paul D. ; Shoji, Mikio ; Kikuchi, Yuichiro ; Yukitake, Hideharu ; Oohara, Naoya ; Naito, Mariko ; Okamoto, Kuniaki ; Reynolds, Eric C. ; Nakayama, Koji. / Identification of a new membrane-associated protein that influences transport/maturation of gingipains and adhesins of Porphyromonas gingivalis. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 10. pp. 8668-8677.
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AU - Sato, Keiko

AU - Sakai, Eiko

AU - Veith, Paul D.

AU - Shoji, Mikio

AU - Kikuchi, Yuichiro

AU - Yukitake, Hideharu

AU - Oohara, Naoya

AU - Naito, Mariko

AU - Okamoto, Kuniaki

AU - Reynolds, Eric C.

AU - Nakayama, Koji

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