| Project/Area Number |
09557175
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
矯正・小児・社会系歯学
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| Research Institution | Osaka University |
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Principal Investigator |
SHIZUKUISHI Satoshi Osaka University, Faculty of Dentistry, Professor, 歯学部, 教授 (00028789)
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| Co-Investigator(Kenkyū-buntansha) |
KATAOKA Kosuke Osaka University, Faculty of Dentistry, Research assosiate, 歯学部, 助手 (50283792)
NAGATA Hideki Osaka University, Faculty of Dentistry, Assistant Professor, 歯学部・附属病院, 講師 (50260641)
AMANO Atsuo Osaka University, Faculty of Dentistry, Assistant Professor, 歯学部・附属病院, 講師 (50193024)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | periodontopathic bacteria / Porphyromonas gingivalis / fimbriae / coaggregation / Streptococcus oralis / proline-rich protein / statherin / binding |
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| Research Abstract |
We previously showed that P.gingivalis fimbriae specifically bind to salivary acidic proline-rich proteins (PRP) and that the active domain in PRP is PQGPPQ.In this study, we examined the effect of synthetic peptide (PQGPPPQGGRPQGPPQGQSPQ ; pepPRP-C) corresponding to C-terminal region in PRP on the interaction between fimbriae and salivary proteins. The pepPRP-C significantly inhibited the binding of fimbriae to basic proline-rich glycoproteins (PRG)-coated hydroxyapatite (HAP) beads as well as to PRP on HAP beads. In the overlay assay, the pepPRP-C clearly diminished the interactions between the fimbriae and the various salivary components, including PRPs, PRGs, and the components with smaller molecular sizes. Moreover, pepPRP-C inhibited the coaggregation of P.gingivalis with various streptococcal strains, suggesting that this peptide may act as an effective inhibitor against P.gingivalis adherence to dental plaque. Then the recombinant Streptococcus gordonii secreting pepPRP-C was generated as a model of a possible approach to prevent the oral colonization of P.gingivalis. A duplicate DNA fragment (pepPRP-C) encoding pepPRP-C was obtained by self-complementary annealing of synthetic oligonucleotides. PepPRP-C was connected downstream to a promoter and a gene encoding a signal peptide of Streptococcus downei glucosyltransferase 1 in frame. The linked fragments were inserted into the plasmid pMNK-4 derived from pVA838. The constructed plasmid was transformed to S.gordonii G9B, which successfully secreted the recombinant pepPRP-C (r-pepPRP-C). The concentrated bacterial culture supernatant containing r-pepPRP-C inhibited the binding of P.gingivalis cells and fimbriae to PRP up to 72% and 77%, respectively. The r-pepPRP-C concentrate also inhibited the coaggregation of P.gingivaLis with various streptococcal strains. Therefore, the pepPR? -C secretion system might be available for prevention against P.gingivalis-induced periodontitis.
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