Haemoglobin receptor protein is intragenically encoded by the cysteine proteinase‐encoding genes and the haemagglutinin‐encoding gene of <i>Porphyromonas gingivalis</i>

Nakayama, Koji; Ratnayake, Dinath B.; Tsukuba, Takayuki; Kadowaki, Tomoko; Yamamoto, Kazuo; Fujimura, Setsuo

doi:10.1046/j.1365-2958.1998.00656.x

Cited by 118 publications

(171 citation statements)

References 52 publications

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“…The adhesin domains consist of HGP44, HGP15, HGP17 and HGP27. HGP44 and HGP17 are believed to be involved in haemagglutination, while HGP15 has the ability to bind haemoglobin (Curtis et al, 1996;Kelly et al, 1997;Nakayama et al, 1998;Shi et al, 1999;Shibata et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

“…Rgp and Kgp activities cause not only destruction of periodontal tissue but also disruption of host-defence mechanisms (Wingrove et al, 1992;Kadowaki et al, 1994;Imamura et al, 1995Imamura et al, , 1997Okamoto et al, 1996;Abe et al, 1998;Calkins et al, 1998;Scragg et al, 1999). In addition, Rgp is involved in fimbriation, by processing the fimbrial subunit protein (FimA) to a mature form Kadowaki et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia

et al. 2003

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Co-aggregation among bacterial cells caused by the adherence of one bacterial species to another is a potential colonization mechanism. Several putative aggregation factors for co-aggregation between Porphyromonas (Por.) gingivalis and Prevotella (Pre.) intermedia were partially purified from Por. gingivalis vesicles by gel filtration and affinity chromatography. Antisera against the aggregation factors were made. Analysis using these antisera revealed that 18 and 44 kDa proteins might be responsible for Por. gingivalis vesicle-mediated aggregation of Pre. intermedia. Using antiserum against the 18 kDa protein, the DNA region encoding it was cloned from Por. gingivalis genomic DNA. Sequence analysis revealed that the DNA region was located within the rgpA and kgp genes, encoding Arg-gingipain (Rgp) and Lys-gingipain (Kgp), respectively, and it encoded non-catalytic adhesin domain regions, namely a C-terminal portion of HGP15, the entire HGP17 sequence and an N-terminal portion of HGP27. A portion of the DNA sequence was also found in the haemagglutinin A (hagA) gene. A recombinant glutathione S-transferase (GST)-HGP17 fusion protein reacted to antiserum against the 18 kDa protein and Pre. intermedia cells could adhere to GST-HGP17-conjugated Sepharose 4B beads, indicating that the HGP17 domain protein is responsible for Por. gingivalis vesicle-mediated aggregation of Pre. intermedia.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia

et al. 2003

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“…Support for this hypothesis has emerged from the work of Lewis et al (1999), which demonstrates that this enzyme is an efficient haemoglobinase and hence may serve to release protein-bound haemin into the extracellular environment. Furthermore, Nakayama et al (1998) have shown that a haemin-binding peptide is intragenically encoded by kgp and this may function as part of a haemin storage mechanism at the cell surface. However, data in this report demonstrate that KGP is not required for growth in complex medium with added haemin.…”

Section: Discussionmentioning

confidence: 99%

Generation of Lys-gingipain protease activity in Porphyromonas gingivalis W50 is independent of Arg-gingipain protease activities

et al. 2000

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Porphyromonas gingivalis, a black-pigmenting anaerobe implicated in the aetiology of periodontal disease, contains two loci, rgpA and rgpB, encoding the extracellular Arg-X specific proteases (RGPs, Arg-gingipains), and kgp, which encodes a Lys-X specific protease (KGP, Lys-gingipain). The rgpA and kgp genes encode polyproteins comprising pro-peptide and catalytic domain with large N-and C-terminal extensions which require proteolytic processing at several Arg and Lys residues to generate mature enzymes. The product of rgpB contains only a pro-peptide and the catalytic domain which requires processing at an Arg residue to generate active enzyme. An rgpA rgpB double mutant (E8) of P. gingivalis was constructed to study the role of RGPs in the processing of KGP. A kgp mutant (K1A) was also studied to investigate the role of KGP in the generation of RGPs. E8 was stable in the absence of the antibiotics tetracycline and clindamycin (selection markers for rgpA and rgpB, respectively) and exhibited the same pigmentation, colony morphology and identical growth rates to the parent W50 strain in the absence of antibiotics, in both complex and chemically defined media. The KGP activity of E8, grown in the absence of tetracycline, in whole cultures and in culture supernatants (up to 6 d) was identical to levels in W50. However, in the presence of tetracycline in the growth medium, the level of KGP was reduced to 50 % of levels present in whole cultures of W50. Since tetracycline had no effect on RGP or KGP activity when incorporated into assay buffer, this effect is most likely to be on the synthesis of Kgp polypeptide. K1A was also stable in the absence of antibiotics but was unable to pigment, and remained straw-coloured throughout growth. RGP activity in whole cultures of K1A was identical to levels in W50, but RGP activity in 6 d culture supernatants was reduced to 50 % of levels present in W50. Thus, although KGP is not required for generation of RGP activity from RgpA and RgpB polypeptides, its absence affects the release/transport of RGP into culture supernatant.

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“…After three 10-min washes with PBS, peroxidase activity was detected (29). For antibody binding, rabbit anti-HbR antiserum (18) and mouse monoclonal antibody (mAb) 61BG1.3 for the detection of the non-HbR domain proteins (30) were used as the primary antibody, and HRP-conjugated anti-rabbit and anti-mouse IgGs were used as the secondary antibody, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…The Cterminal adhesin domain region of rgpA consists of four domains (HGP44, HGP15, HGP17, and HGP27) (15). One of the domain proteins, HGP15, was found to have the ability to bind hemoglobin by surface plasmon resonance detection using a recombinant HGP15 protein, and we proposed to designate this protein "hemoglobin receptor (HbR) domain protein" (18). The three other non-HbR domains (HGP44, HGP17, and HGP27) have a 49-amino acid-long sequence in common (15).…”

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confidence: 99%

Genetic Analyses of Proteolysis, Hemoglobin Binding, and Hemagglutination of Porphyromonas gingivalis

Shi¹,

Ratnayake²,

Okamoto³

et al. 1999

Journal of Biological Chemistry

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Porphyromonas gingivalis produces arginine-specific cysteine proteinase (Arg-gingipain, RGP) and lysinespecific cysteine proteinase (Lys-gingipain, KGP) in the extracellular and cell-associated forms. Two separate genes (rgpA and rgpB) and a single gene (kgp) have been found to encode RGP and KGP, respectively. We constructed rgpA rgpB kgp triple mutants by homologous recombination with cloned rgp and kgp DNA interrupted by drug resistance gene markers. The triple mutants showed no RGP or KGP activity in either cell extracts or culture supernatants. The culture supernatants of the triple mutants grown in a rich medium had no proteolytic activity toward bovine serum albumin or gelatin derived from human type I collagen. Moreover, the mutants did not grow in a defined medium containing bovine serum albumin as the sole carbon/energy source. These results indicate that the proteolytic activity of P. gingivalis toward bovine serum albumin and gelatin derived from human type I collagen appears to be attributable to RGP and KGP. The hemagglutinin gene hagA of P. gingivalis possesses the adhesin domain regions responsible for hemagglutination and hemoglobin binding that are also located in the Cterminal regions of rgpA and kgp. A rgpA kgp hagA triple mutant constructed in this study exhibited no hemagglutination using sheep erythrocytes or hemoglobin binding activity, as determined by a solid-phase binding assay with horseradish peroxidase-conjugated human hemoglobin, indicating that the adhesin domains seem to be particularly important for P. gingivalis cells to agglutinate erythrocytes and bind hemoglobin, leading to heme acquisition.

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Haemoglobin receptor protein is intragenically encoded by the cysteine proteinase‐encoding genes and the haemagglutinin‐encoding gene of Porphyromonas gingivalis

Cited by 118 publications

References 52 publications

Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia

Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia

Generation of Lys-gingipain protease activity in Porphyromonas gingivalis W50 is independent of Arg-gingipain protease activities

Genetic Analyses of Proteolysis, Hemoglobin Binding, and Hemagglutination of Porphyromonas gingivalis

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