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Epitope structure of fibronectin that recognized by bacterial adhesins

Research Project

Project/Area Number 11671873
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 病態科学系歯学(含放射線系歯学)
Research InstitutionKagoshima University

Principal Investigator

ITO Hiro-o  Kagoshima University, Dental School, Associate Professor, 歯学部, 助教授 (40213079)

Co-Investigator(Kenkyū-buntansha) SOUTOME Sakiko  Kagoshima University, Dental School, Research Associate, 歯学部, 助手 (20325799)
KITADA Katsuhiro  Kagoshima University, Dental School, Research Associate, 歯学部, 助手 (90195264)
岡田 康子  鹿児島大学, 歯学部, 助手 (90315424)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1999: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsFibronectin / Extracellular Matrix Protein / Infective Endocarditis / Monoclonal Antibodies / Epitope Mapping / Viridans Streptococci / Oral Indioenous Bacteria / 口腔内細菌 / 常在細菌 / 細胞外基質タンパク / エピトープ
Research Abstract

Fibronectin (Fn) is a major extracellular matrix protein and appears to mediate adherence of various microorganisms to host tissues at the first stage of infections. Granulicatella adiacens, a dominant nutritionally variant streptococci (NVS) harboring in the healthy human oral cavities, is often isolated from patients with infective endocarditis as the causative agent. We have previously reported a link between the Fn-binding ability of this specie and the infectivity in an animal model of endocarditis. We report herein that Fn possesses a novel binding site for various species of bacteria including this organism, and the region may be more relevant to microbial pathogenesis than the N-terminal region which has been extensively investigated from this point of view. Monoclonal antibodies (mAbs) were generated against human Fn, and one clone with an ability to inhibit the Fn-binding of G. adiacens was selected. Western blotting analyses using polypeptide fragments of Fn obtained by sequ … More ential limited digestions with thermolysin and lysyl endopeptidase showed that the epitope for the inhibiting mAb resided in the central part of the whole Fn polypeptide (105 kDa cell-binding fragment) which lacked both the N- and C-terminal parts, the former contains the known binding regions for many bacterial species. Staphylococcus aureus and Streptococcus pyogenes were also capable of binding to the central 105 kDa cell-binding fragment immobilized onto assay plates, in addition to the known binding to the N-terminal part while G. adiacens and an Echerichia coli were only to the central part. The adherence of S. aureus and S. pyogenes to N-terminal Fn fragment was inhibited by excess amounts of free Fn in the assay medium. In contrast, adherence of the four species to the central 105 kDa fragment was not inhibited by soluble Fn, but by the mAb. Considering the high concentrations of soluble Fn in the mammalian body fluids, the central part of Fn may have more important roles than the N-terminal region in the bacterial adherence in vivo. Less

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] Ito, H.-O.et al.: "Inhibition of fibronectin-binding of some bacterial cells by subtle pH increase within the physiological range"J. Microbiol. Meth.. 53(印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ito, H.-O., Soutome, S., Inoue, M.: "Inhibition of fibronectin-binding of some bacterial cells by subtle pH increase within the physiological range"J. Microbiol. Meth.. (in press). (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 五月女さき子: "Abiotrophia adiacensのフィブロネクチン付着を阻害するモノクローナル抗体の作製"口腔衛生会誌. 50・4. 590-591 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Soutome,S.: "Identification of a New Bacterial Binding Region of Fibronectin Molecule"J.Dent.Res. (in press).

    • Related Report
      2000 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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