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1991 Fiscal Year Final Research Report Summary

Analysis of Resistance Mechanism of Complement-resistant Bacteria

Research Project

Project/Area Number 01480178
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionOsaka University

Principal Investigator

INOUE Kozo  Osaka Univ. Med. Sch., Professor, 医学部, 教授 (10028300)

Co-Investigator(Kenkyū-buntansha) INAGI Reiko  Osaka Univ. Med. Sch., Research Associate, 医学部, 助手
HONG Kyongsu  Osaka Univ. Med. Sch., Research Associate, 医学部, 助手 (30158894)
Project Period (FY) 1989 – 1991
KeywordsSerum-resistant Bacteria / Phagocytosis / Bactericidal Activity / Staphylococcal M Protein / traT gene of R factor
Research Abstract

1. Streptococcal M protein inhibited significantly the alternative pathway C3 convertase of complement and the classical C5 convertase. Therefore, M protein intercepts the amplification circuit of C3b formation and also inhibits the formation of C5a, leading to suppressed expression of complement receptors (CR1 and CR3) on the phagocytic cells.
2. A murine monoclonal antibody was raised against the synthetic TraT peptide (86-99). This MoAb reated with only denatured TraT protein. Using this MoAb for monotoring TraT by immunoblotting, TraT protein was partially purified from solubilized membrane fraction of traT^+ bacteria. Six MoAbs were then generated against this protein. These MoAbs reacted with the native protein even on the living bacteria, and their Fab fragments suppressed the inhibiting effect of TraT protein on bactericidal activity of serum.
The purification method was established from solubilized bacterial membrane fraction by ion exchange and gel filtration chromatographies. The final preparation contained no endotoxin activity.
The purified TraT protein inhibited the lysis of sensitized erythrocytes by serum complement. Its inhibitory action was mainly on the C6 step. It also slightly inhibited the reaction steps of C4, C5 and factor B. However, TraT inhibited neither the cleavage of C5 nor the reaction of preformed C5b6 complex.
The TraT protein probably inhibits the formation of C5b6 complex or causes formation of structurally alterated, nonfunctional C5b6 complex.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Hong,K.: "Inhibition of the alternative C3 convertase and classical C5 convertase of complement by group A streptococcal M protein." Infection and Immunity. 58. 2535-2541 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kozono,H.: "Localization of the covalent C3b binding site on C4b within the complement classical pathway C5 convertase,C4b2a3b." Journal of Biological Chemistry. 265. 14444-14449 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hong,K.: "Restriction of C5 convertase of the alternative complement pathway with isolated C3b dimer and factors B and D." Journal of Immunology. 146. 1868-1873 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Pramoonjago,P.: "Bactericidal activity of C9-deficient human serum." Journal of Immunology. 148. 837-843 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Pramoonjago,P.: "Role of TraT protein,an anticomplementary protein produced in Escherichia coli by factor R100,in serum resistance." Journal of Immunology. 148. 827-836 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kim,Y.U.: "Covalent binding of C3b to C4b within the classical complement pathway C5 convertase.Determination of amino acid residues involved in ester linkage formation." Journal of Biological Chemistry. 267. (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 井上 公藏: "医科細菌学(吉川昌之介編集)「細菌の感染と免疫・非特異的因子」" 南江堂(東京), 325 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 木下 タロウ,井上 公藏: "医学細菌学(三輪谷俊夫監修)Vol.5.「感染防御における補体の役割」" 菜根出版(東京), 323 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hong, K., T. Kinoshita, J. Takada, P. Pramoonjage, Y. U. Kim, and K. Inoue: "Inhibition of the alternative C3 convertase and classical C5 convertase of complement by group A streptococcal M protein." Infect. Immun. 58. 2535-2541 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Pramoonjago, P., M. Kaneko, T. Kinoshita, E. Otsubo, J. Takeda, K. Hong, R. Inagi, and K. Inoue.: "Role of TraT protein, an anticomplementary protein produced in Escherichia coli by R100 factor, in serum resistance." J. Immunol.148. 827-86 (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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