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MECHANISM OF ACTION OF PORE FORMING TOXINS

Research Project

Project/Area Number 06454206
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

SHINODA Sumio  Okayama University, Faculty of Pharmaceutical Sciences Professor, 薬学部, 教授 (50029782)

Co-Investigator(Kenkyū-buntansha) MIYOSHI Shin-ichi  Okayama University, Faculty of Pharmaceutical Sciences Research Associate, 自然科学研究科, 助手 (60182060)
TOMOCHIKA Ken-ichi  Okayama University, Faculty of Pharmaceutical Sciences Associate Professor, 薬学部, 助教授 (00093691)
KATSU Takashi  Okayama University, Faculty of Pharmaceutical Sciences Associate Professor, 薬学部, 助教授 (40112156)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1994: ¥4,200,000 (Direct Cost: ¥4,200,000)
KeywordsPathogenic vibrios / Hemolysin / Osmotic hemolysis / Vibrio mimicus / Vibrio vulnificus / Vibrio fluvialis / PCR
Research Abstract

Bacteria of genus Vibrio are normal habitant in natural environmental water. Because of universal consumption of sea foods in Japan, gastroenteric infections due to pathogenic vibrios break out frequently, therefore it is an important problem to clarify the mechanism of infection of the vibrios. Many pathogenic vibrios produce hemolysin as an important pathogenic factor. It is expected that elucidation of action mechanism of the hemolysins leads to development of methods for prevention and therapy of the vibrio infections. We selected two vibrio hemolysins, Vibrio vulnificus hemolysin (VVH) and V.mimicus hemolysin (VMH), and carried out the comparative study of action mechanism in vitro and in vivo. The project started in 1994. Differences of properties of the hemolysins produced by different strains and a role of VMH as a diarrhenogenic factor were demonstrated in 1994 and 1995. In 1996, protein-chemical and immuno-chemical studies of VMH,especially structure and function, were carried out. VMH was purified with ethylene glycol as the solvent. Monoclonal antibodies (MAb) against VMH were prepared but only IgM MAbs which had binding activity to VMH but not neutralizing activity were obtained. Furthermore, non-immunized IgMs also showed weak binding activity to VMH which was suspected to bind to sialic acid on the IgM molecule. The IgM antibodies, however, showed neutralizing activity to VMH purified by a modified method without ethylene glycol, suggesting the conformational change of recognition site for sialic acid of the membrane surface by ethylene glycol, although it is not so distinguish detectable by physico-chemical methods such as fluorescence analysis. Informations on mechanism of binding of VMH molecule to target cells obtained in this project are useful for understanding of the mechanism of gastroenteritis due to the vibrio.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Oh,E.-G.: "Homogeneity of hemolysin produced by Vibrio vulnificus isoates" Journal of Natural Toxins. 3. 177-123 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam,M.: "Production of antigenically related exocellular elastolytic proteases mediating hemagglutination by vibrios" Microbiology and Immunology. 38. 67-70 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam,M.: "Expression of virulence-related properties by,and intestinal adhessiveness of,Vibrio mimicus strains isolated from aquatic environments." Applied and Environmental Microbiology. 62. 3871-3874 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam,M.: "Purification and characterization of novel hemagglutinins from Vibrio mimicus : a 39 kDa major outer membrane protein and lipopolysaccharide" Infection and Immunity. 64. 4035-4041 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Rahman,M.M.: "Analysis of the structural gene encoding a hemolysin in Vibrio mimicus" Microbiology and Immunology. 41. 169-173 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miyoshi,S.: "Purification and characterization of a hemolysin produced by Vibrio mimicus" Infection and Immunity. 65(印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Oh, E.-G., Yamanaka, H., Ohmae, K., Kim, Y.-M., Usui, K., Miyoshi, S., Shinoda, S.: "Homogeneity of hemolysin produced by Vibrio vulnificus isolates." J.Natural Toxins. 3. 117-123 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam, M., Miyoshi, S., Shinoda, S.: "Production of antigenically related exocellular elastolytic proteases mediating hemagglutination by vibrios." Microbiol.Immunol.38. 67-70 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam, M., Miyoshi, S., Yamamoto, S., Tomochika, K., Shinoda, S.: "Expression of virulence-related properties by, and intestinal adhesiveness of, Vibrio mimicus strains isolated from aquatic environments." Appl.Env.Microbiol.62. 3871-3874 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam, M., Miyoshi, S., Tomochika, K., Shinoda, S.: "Purification and characterization of novel hemagglutinins from Vibrio mimicus : a 39 kDa major outer membrane protein and lipopolysaccharide." Infect.Immun.64. 4035-4041 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Rahman, M.M., Miyoshi, S., Tomochika, K., Wakae, H., Shinoda, S.: "Analysis of the structural gene encoding a hemolysin in Vibrio mimicus." Microbiol.Immunol.41. 169-173 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miyoshi, S., Sasahara, K., Akamatsu, S., Rahman, M.M., Katsu, T., Tomochika, K., Shinoda, S.: "Purification and characterization of a hemolysin produced by Vibrio mimicus." Infect.Immun.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Alam,M.,Miyoshi,S.,Yamamoto,S.,Tomochika,K.,Shinoda,S.: "Expression of virulence-related properties by,and intestinal adhessiveness of,Vibrio mimicus strains isolated from aquatic environments." Applied and Environmental Microbiology. 62. 3871-3874 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Alam,M.,Miyoshi,S.,Tomochika,K.,Shinoda,S.: "Purification and characterization of novel hemagglutinins from Vibrio mimicus : a 39 kDa major outer membrane protein and lipolysaccharide." Infection and Immunity. 64. 4035-4041 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Rahman,M.,Miyoshi,S.,Wakae,H.,Tmochika,K.,Shinoda,S.: "Analysis of the structural gene encoding a hemolysin in Vibrio mimicus." Microbiology and Immunology. 41. 169-173 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Munirul Alam et al.: "Production of Antigenically Related Exocellular Elastolytic Proteases Mediating Hemagglutination by Vibrios" Microbiology and Immunology. 39. 67-70 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yasuhiro Shimonaka et al.: "Changes in the Shape of Escherichia coli and Human Erythrocytes induced by Antimicrobial Peptides : Polymyxin B,EM 49,Gramicidin S and Tachyplesin" Peptide Chemistry 1994. 69-72 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Eun-Gyoung Oh: "HOMOGENEITY OF HEMOLYSIN PRODUCED BY VIBRIO VULNIFICUS ISOLATES" Journal of Natural Toxins. 3. 117-123 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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