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Characterization of Heat Shock Proteins from Periodontopathogenic Bacteria and Their Virulences

Research Project

Project/Area Number 08672372
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 矯正・小児・社会系歯学
Research InstitutionThe University of Tokushima

Principal Investigator

HINODE Daisuke  The University of Tokushima, School of Dentistry, Associate Professor, 歯学部, 助教授 (70189801)

Co-Investigator(Kenkyū-buntansha) TANABE Shin-ichi  The University of Tokushima, University Dental Hospital, Research Associate, 歯学部・附属病院, 助手 (40284301)
OTSUKA Chiaki  The University of Tokushima, School of Dentistry, Research Associate, 歯学部, 助手 (00263848)
TAMATANI Kanako  The University of Tokushima, School of Dentistry, Research Associate, 歯学部, 助手 (40243711)
NAKAMURA Ryo  The University of Tokushima, School of Dentistry, Professor, 歯学部, 教授 (30034169)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsHeat shock protein / Periodontopathogenic bacteria / Immunological characterization / Purification / Specific antibody / Bacteroides forsythus / Porphromonas gingivalis / Actinobacillus actinomycetemcomitans / 熱ショックタンパク質 / 歯周病原細菌 / 歯周病原性
Research Abstract

Recently, considerable attention has been given to the potential roles of heat shock protein (HSP) of in inflammation and autoimmune diseases. In this study, we established the isolation procedure of HSP from periodontopathogenic bacteria and investigated the immunological characterization of them using the specific antibodies.
The isolation procedure of HSPs from whole cells of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Bacteroides forsythus was performed by affinity chromatography on adenosine 5'-triphosphate-agarose followed by preparative polyacrylmide gel electrophoresis and could obtain the considerable amount of purified GroEL- and DnaK-like proteins. Also, these proteins could be successfully used to obtain rabbit sera which possessed high titer of antibodies and a very good specificities comparing to the commercial ones.
By Western-immunoblotting, it found that the GroEL-like protein from P.gingivalis was localized in cytoplasm and the DnaK-like protein f … More rom P.gingivalis was localized in both cytoplasm and periplasm.
Analysis of the N-terminal amino acid sequence of each DnaK-like protein from P.gingivalis and B.forsythus showed a high degree of homology with the DnaK protein from Escherichia coli.
However, these two HSPs reacted very weakly with a commercial anti-DnaK polyclonal antibody by dot-blotting.
GroEL-like proteins isolated P.gingivalis, A.actinomycetemcomitans and B.forsythus showed a high degree of homology of their N-terminal amino acid sequences. Polyclonal antibodies raised against each GroEL-like protein showed a high level of cross-reactivity. Reactivity of these antibodies against recombinant human HSP60 was weak.
Our findings suggest that DnaK- and GroEL-like proteins from periodontopathogens are well conserved and that the GroEL-like protein resemble each other more closely. It is possible to suggest that HSPs from bacteria are important targets of the immune response, however, these anti-infectious immune response may shift from their own protection to pathogenesis by their molecular mimicry. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Hinode, Daisuke et al.: "A general procedure for the isolation of heat shock proteins from periodontopathogenic bacteria" J.Microbiol. Methods. 25. 349-355 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 日野出 大輔、他5名: "歯周病原性細菌由来のDnaK様蛋白質の免疫学的研究" 口腔衛生学会雑誌. 46. 532-533 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 日野出 大輔、他5名: "Prophyromonas gingivalisの熱ショック蛋白質" 口腔衛生学会雑誌. 47. 412-413 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hinode, Daisuke et al.: "Cross-reactivity of Specific antibodies directed to that shock proteins from periodontopathogenic bacteria and of human origin" Oral Microbiol.Immunol.13. 55-58 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hinode, D.et al.: "A general procedure for the isolation of heat shock proteins from periodontopathogenic bacteria" J.Microbiol.Methods. 25. 349-355 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hinode, D.et al.: "Immunological studies of the DnaK-like protein from periodontopathogenic bacteria" J.Dental Health. 46. 532-533 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hinode, D.et al.: "Heat shock proteins from Porphyromonas gingivalis" J.Dental Health. 47. 412-413 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hinode.D.et al.: "Cross-reactivity of Specific antibodies directed to heat shock proteins from periodontopathogenic bacteria and of human origin" Oral Microbiol.Immunol.13. 55-58 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 日野出大輔、他5名: "歯周病原性細菌由来のDnaK様蛋白質の免疫学的研究" 口腔衛生学会雑誌. 46. 532-533 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 日野出大輔、他5名: "Porphyromonas gingivalis の熱ショック蛋白質" 口腔衛生学会雑誌. 47. 412-413 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hinode,Daisuke et al.: "Cross-reactivity of Specific antibodies directed to heat shock proteins from periodontopathogenic bacteria and of human origin" oral Microbiol.Immunol.13. 55-58 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hinode,D.,Grenier,D.and Mayrand,D.: "A general procedure for the isolation of heat-shock proteins from periodontopathogenic bacteria" Journal of Microbiological Methods. 25. 349-355 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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