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Analysis of toxic region of S. hyicus exfoliative toxin using mutant proteins

Research Project

Project/Area Number 13660307
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Basic veterinary science/Basic zootechnical science
Research InstitutionKitasato University

Principal Investigator

MAEHARA Nobutoshi  Kitasato Univ., Sch. Vet. Med. & Anim. Sci., Professor, 獣医畜産学部, 教授 (90072371)

Co-Investigator(Kenkyū-buntansha) SATO Hisaaki  Kitasato Univ., Sch. Vet. Med. & Anim. Sci., Assistant Professor, 獣医畜産学部, 講師 (40154083)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsS. hyicus / Exfoliative toxin / Mutant protein / Active region
Research Abstract

To analyze the active region of Staphylococcus hyicus exfoliative toxin B (SHETB), mutants of His^<87> (H87A, in which His^<87> was mutated to Ala), Asp^<135> (D135A, in which Asp^<135> was mutated to Ala), Ser^<209> (S209C, in which Ser^<209> was mutated to Cys), Lys^<227> (K227E, in which Lys^<227> was mutated to Glu) and Tyr^<180> (Y180F, in which Tyr^<180> was mutated to Phe), were synthesized by site-directed mutagenesis method.
When SHETB and five mutants were inoculated to NCTC2544 cells (SHETB-susceptible cell lines) and incubated at 37℃ for 48 h, the rounding effect was appeared on the cells inoculated with SHETB, K227E and Y180F. However the rounding were not appeared on the cells inoculated with H87A, D135A and S209C. The exfoliation (Nikolsky sign) was seen in the 1-day-old chicks (SHETB-susceptible animal) inoculated with SHETB, K227E and Y180F, but not seen in the chicks inoculated with H87A, D135A and S209C.
These findings suggest that active region of SHETB is constructed by His^<87>, Asp^<135> and Ser^<209>. The regions, in which included in the above three residues, is conservative among the various serine proteases. Therefore, the toxic activity of SHETB seems to be proteolytic activity of serine protease.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 佐藤久聡: "ブドウ球菌表皮剥脱毒素の構造と活性"獣医生化学. 38・1. 1-11 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terauchi, R., Sato, H., Maehara, N.et al.: "Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs"Veterinary Microbiology. (in print). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terauchi, R., Sato, H., Maehara, N.et al.: "Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli"Veterinary Microbiology. (in print). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sato, H.: "Structure and activity of staphylococcal exfoliative toxins (in Japanese with English abstract)"Veterinary Biochemistry. 38-1. 1-11 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terauchi, R., Sato, H., Maehara, N. et al.: "Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs"Veterinary Microbiology. in print. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Terauchi, R., Sato, H., Maehara, N. et al.: "Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli"Veterinary Microbiology. in print. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 佐藤久聡: "ブドウ球菌表皮剥脱毒素の構造と活性"獣医生化学. 38・1. 1-11 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] Terauchi, R., Sato, H., Maehara, N.et al.: "Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs"Veterinary Microbiology. (in print). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Terauchi, R., Sato, H., Maehara, N.et al.: "Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli"Veterinary Microbiology. (in print). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 佐藤 久聡: "ブドウ球菌表皮剥脱毒素の構造と活性"獣医生化学. 38・1. 1-11 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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