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Mechnism of internalization of Clostridium botulinum toxin through the neural membrane

Research Project

Project/Area Number 60560334
Research Category

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

Allocation TypeSingle-year Grants
Research Field Applied veterinary science
Research InstitutionUniversity of Osaka Prefecture

Principal Investigator

SAKAGUCHI Genji  University of Osaka Prefecture, College of Agriculture, 農学部, 教授 (50081477)

Co-Investigator(Kenkyū-buntansha) NISHIMURA Masakazu  University of Osaka Prefecture, College of Agriculture, 農学部, 助教授 (50011995)
KAMATA Yoichi  University of Osaka Prefecture, College of Agriculture, 農学部, 助手 (20152837)
KOZAKI Shunji  University of Osaka Prefecture, College of Agriculture, 農学部, 助手 (10109895)
Project Period (FY) 1985 – 1986
Project Status Completed (Fiscal Year 1986)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1986: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1985: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsClostridium botulinum / Neurotoxin / Monoclonal antibody / Acetylcholine / フラグメント
Research Abstract

Clostridium botulinum neurotoxin types A through G acts presynaptically on nerve endings to inhibit acetylcholine release. The neurotoxin is composed of two fragments, the heavy chain(H) of 100K and the light chain(L) of 50K. For the toxic action, these two fragments function complementarily. We analyzed the fine antigenic structure and the function of the neurotoxin with monoclonal antibodies and demonstrated that the neurotoxin molecule comprises at least three fragments named L, H-1 and H-2. The site of binding to the brain synaptosomal membrane was located on H-2, which is located at the C-terminus of the neurotoxin molecule. The structure of the ganglioside the toxin binds, which is regarded as a receptor candidate, was located on domains differed from one type to another. Type E neurotoxin bound to ganglioside <G_(Q1b)> , whereas type B hardly did so. L・H-1 fragment lacking H-2 did not bind to any ganglioside, but to cerebroside and free fatty acids composing gangliosides. These … More results suggest that H-2 binds to the sugar chain, forming the hydrophilic portion of ganglioside, and that L・H-1 fragment, particularly H-1 binds to the hydrophobic portion. H-2 binds to the receptor on the surface of the neural membrane and H-1 plays an important role in the toxin internalization. By use of mouse antineurotoxin antibody and gold colloid-labelled anti-mouse IgG, toxin molecules were located under an electron microscope at the mouse myoneural junction of the mice exposed to the toxin in vivo and in vitro. Gold colloid was detected on the external membrane of the myoneural junction, invaginated parts of the membrane, in the synaptic vesicles, and the cytoplasm of the ending. The toxin translocates into the nerve endings in association with the formation of synaptic vesicles, followed by internalization into the cytoplasm. Further studies are in progress to correlate the function of each fragment of the toxin found by analysis with monoclonal antibodies and the localization of the toxin at the nerve endings. Less

Report

(1 results)
  • 1986 Final Research Report Summary
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Sakaguchi,Genji: FEMS Microbiology Letters. 33. 23-29 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Kozaki,Shunji: Infection and Immunity. 52. 786-791 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Kamata,Yoichi: The Japanese Journal of Veterinary Science. 48. 909-914 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Takamizawa,Kotaro: FEBS Letters. 201. 229-232 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Kamata,Yoichi: Biochemical and Biophysical Research Communications. 140. 1015-1019 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] F.Dorner and J.Drews,eds.Sakaguchi,Genji: "Clostridium botulinum toxins.Pharmacology of Bacterial Toxins.International Encyclopedia of Pharmacology and Therapeuticcs.Section 119." Pergamon Press,Oxford,England, (519-548) (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] SAKAGUCHI, Genji: "Purification and some properties of Clostridium botulinum type AB toxin" FEMS Microbiology Letters. 33. 23-29 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] KOZAKI, Shunji: "The use of monoclonal antibodies to analyze the structure of Clostridium botulinum type E derivative toxin" Infection and Immunity. 52. 786-791 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] KAMATA, Yoichi: "Evaluation of ELISA techniques for titration of monoclonal antibodies against botulinum toxin" Japanese Journal of Veterinary Science. 48. 909-914 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] TAKAMIZAWA, Kotaro: "TLC immunostaining characterization of Clostridium botulinum type A neurotoxin binding to gangliosides and free fatty acids." FEBS Letters. 201. 229-232 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] KAMATA,Yoichi: "Evidence for direct binding of Clostridium botulinum type E derivative toxin and its fragments to gangliosides and free fatty acids" Biochemical and Biophysical Research Communication. 140. 1015-1019 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] SAKAGUCHI, Genji: Pergamon Press, Oxford, England. Clostridium botulinum toxins. p. 519-548. In Pharmacology of Bacterial Toxins. International Encyclopedia of Pharmacology and Therapeutics. Section 119 [F. Dorner and J. Drews (eds)]., 748 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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