Molecular cloning and characterization of the genes encoding the immunoreactive major cell-surface proteins of Porphyromonas gingivalis

Ogawa, Teiichiro

doi:10.1016/0378-1097(94)00170-7

Cited by 11 publications

(25 citation statements)

References 12 publications

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“…Although a search was conducted to identify the cleavage sites (33), there were no homologous or similar sequences known as bacterial signal peptide cleavage sequences; at the -3 residue of Ala, Gly, Ser, Val or Ile, and at the -1 residue of Ala, Gly or Ser relative to the cleavage site (42). Furthermore, there were no such cleavage sequences of Arg (-1) -Ala (+1) and -3 residue of Ser found in the cases of fimA (31) or for the gene of the 75 kDa protein, an outer membrane protein (30), in this organism.…”

Section: Discussionmentioning

confidence: 94%

“…Since then other P gingivalis genes have been sequenced, including those for superoxide dismutase (4,28), collagenase (17), a surface protein (30), proteases (3,9,32) and hemagglutinin (Progulske-Fox, A. et al, GenBank accession number Z35494). However, none of the extensive studies regarding genes or gene structures of complex cellular components such as fimbriae have been reported on this organism.…”

Section: Discussionmentioning

confidence: 99%

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Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene (fimA) of the Oral Anaerobe Porphyromonas gingivalis

Watanabe

Onoe

Ozeki³

et al. 1996

Microbiology and Immunology

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The downstream DNA region of the fimbrilin gene (fimA), which encodes the major subunit protein of Porphyromonas gingivalis fimbriae, was fully sequenced. Gene products, expressed from this region in Escherichia coli, were purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their partial amino acid sequences were determined to verify open reading frames (ORFs) found in the region by DNA sequencing. Four ORFs, designated ORF1, ORF2, ORF3 and ORF4, were found in the 5.8-kb PstI fragment downstream from fimA, which was previously cloned and partially characterized by Yoshimura, Takahashi, Hibi, Takasawa, Kato, and Dickinson (Infect. Immun. 61: 5181-5189, 1993). The direction of transcription of all the ORFs was the same as that of fimA. The 50 and 80 kDa encoded proteins, ORF2 and ORF3, respectively, have been reported to be minor components associated with fimbriae. The 15 and 19 kDa proteins, ORF1 and ORF4, respectively, have been expressed in E. coli but not identified in P. gingivalis. However, all the gene products of the ORFs, expressed in E. coli, appeared to contain intact signal peptides based on their N-terminal amino acid sequences.

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene (fimA) of the Oral Anaerobe Porphyromonas gingivalis

Watanabe

Onoe

Ozeki³

et al. 1996

Microbiology and Immunology

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“…The 75-kDa protein, one of the major immunodominant cell surface proteins of P. gingivalis, seems to be processed to a mature form by cleavage between residues Arg-49 and Ala-50 (16,19,32,35). To determine whether RGP is also involved in this maturation, immunoblot analysis of the 75-kDa protein of the RGP mutants was performed (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The 75-kDa protein, one of the highly immunogenic proteins of P. gingivalis, is present in the outer membrane or the outermost part of the organism as a stable, large complex with an apparent molecular mass of 2,000 kDa and seems to contribute to the hostbacterium interaction (35). The precursor form of the 75-kDa protein is also suggested to produce the mature form by specific cleavage between residues Arg-49 and Ala-50 (16,19). Although these cell surface proteins are initially synthesized as the larger precursor proteins and then undergo proteolytic processing during or after translocation to the cell surface through the inner membrane and the periplasm to give rise to the mature forms, it is still unknown what processing enzyme(s) is responsible for the maturation of the proteins.…”

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confidence: 99%

Involvement of arginine-specific cysteine proteinase (Arg-gingipain) in fimbriation of Porphyromonas gingivalis

et al. 1996

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Arginine-specific cysteine proteinase (Arg-gingipain [RGP]), a major proteinase secreted from the oral anaerobic bacterium Porphyromonas gingivalis, is encoded by two separate genes (rgpA and rgpB) on the P. gingivalis chromosome and widely implicated as an important virulence factor in the pathogenesis of periodontal disease (K. Nakayama, T. Kadowaki, K. Okamoto, and K. Yamamoto, J. Biol. Chem. 270:23619-23626, 1995). In this study, we investigated the role of RGP in the formation of P. gingivalis fimbriae which are thought to mediate adhesion of the organism to the oral surface by use of the rgp mutants. Electron microscopic observation revealed that the rgpA rgpB double (RGP-null) mutant possessed very few fimbriae on the cell surface, whereas the number of fimbriae of the rgpA or rgpB mutant was similar to that of the wild-type parent strain. The rgpB ؉ revertants that were isolated from the double mutant and recovered 20 to 40% of RGP activity of the wild-type parent possessed as many fimbriae as the wild-type parent, indicating that RGP significantly contributes to the fimbriation of P. gingivalis as well as to the degradation of various host proteins, disturbance of host defense mechanisms, and hemagglutination. Immunoblot analysis of cell extracts of these mutants with antifimbrilin antiserum revealed that the rgpA rgpB double mutant produced small amounts of two immunoreactive proteins with molecular masses of 45 and 43 kDa, corresponding to those of the precursor and mature forms of fimbrilin, respectively. The result suggests that RGP may function as a processing proteinase for fimbrilin maturation. In addition, a precursor form of the 75-kDa protein, one of the major outer membrane proteins of P. gingivalis, was accumulated in the rgpA rgpB double mutant but not in the single mutants and the revertants, suggesting an extensive role for RGP in the maturation of some of the cell surface proteins.Porphyromonas (Bacteroides) gingivalis, an oral anaerobic bacterium, has been isolated frequently from subgingival lesions in patients with adult periodontal disease, implying that the microorganism is one of the most important pathogens for the disease (25,31,36). P. gingivalis possesses several potential virulence factors (13). Among them, secretory proteinases have received much attention because they can degrade host tissue and cause inflammation (9, 27). Many proteinases have been isolated from culture supernatants, vesicles, membrane fractions, and cell extracts of P. gingivalis (for a review, see reference 5), but it has recently been reported that the proteinases with trypsin-like activity are derived from either Arggingipain (RGP) or Lys-gingipain (24). In a previous study (7), we reported the isolation of RGP (formerly, argingipain) from the culture supernatant of P. gingivalis and its unusual catalytic features related to periodontopathogenicity of the organism (e.g., the abilities to degrade periodontal tissue components, to evade inactivation by internal proteinase inhibitors such as cystatins and ...

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“…Little is known about the morphology of these fimbriae, however, in part because purification is difficult in the presence of the long fimbriae. Nonetheless, it is now well established that the 75-kDa protein, Mfa1 (67 kDa), and PgII (72 kDa) are the same polypeptides, based on their identical N-terminal amino acid sequences and extensive similarity of primary amino acid sequence deduced from the gene sequences (9,26,41). Furthermore, the short fimbriae comprised of Mfa1 are distinct from a third fimbrial type consisting of a 53-kDa protein (1).…”

Section: Vol 73 2005mentioning

confidence: 99%

Short Fimbriae of Porphyromonas gingivalis and Their Role in Coadhesion with Streptococcus gordonii

et al. 2005

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Porphyromonas gingivalis, one of the causative agents of adult periodontitis, attaches and forms biofilms on substrata of Streptococcus gordonii. Coadhesion and biofilm development between these organisms requires the interaction of the short fimbriae of P. gingivalis with the SspB streptococcal surface polypeptide. In this study we investigated the structure and binding activities of the short fimbriae of P. gingivalis. Electron microscopy showed that isolated short fimbriae have an average length of 103 nm and exhibit a helical structure with a pitch of ca. 27 nm. Mfa1, the major protein subunit of the short fimbriae, bound to SspB protein, and this reaction was inhibited by purified recombinant Mfa1 and monospecifc anti-Mfa1 serum in a dose-dependent manner. Complementation of a polar Mfa1 mutant with the mfa1 gene restored the coadhesion phenotype of P. gingivalis. Hence, the Mfa1 structural fimbrial subunit does not require accessory proteins for binding to SspB. Furthermore, the interaction of Mfa1 with SspB is necessary for optimal coadhesion between P. gingivalis and S. gordonii.

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Molecular cloning and characterization of the genes encoding the immunoreactive major cell-surface proteins of Porphyromonas gingivalis

Cited by 11 publications

References 12 publications

Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene (fimA) of the Oral Anaerobe Porphyromonas gingivalis

Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene (fimA) of the Oral Anaerobe Porphyromonas gingivalis

Involvement of arginine-specific cysteine proteinase (Arg-gingipain) in fimbriation of Porphyromonas gingivalis

Short Fimbriae of Porphyromonas gingivalis and Their Role in Coadhesion with Streptococcus gordonii

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