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Analysis fo the role of a novel ovarian protein during oocyte maturation using genetic engineering procedures

Research Project

Project/Area Number 06454682
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Developmental biology
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

TAKAHASHI Takayuki  Hokkaido Univ., Grad.Sch.Sci.Professor, 大学院・理学研究科, 教授 (80197152)

Co-Investigator(Kenkyū-buntansha) OHNISHI Junji  Hokkaido Univ., Grad.Sch.Sci.Instuctor, 大学院・理学研究科, 助手 (40261276)
山下 正兼  北海道大学, 大学院・理学研究科, 助教授 (30202378)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1995: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywordsmammal / ovary / oocyte / ovarian protein / genetic engineering / follipsin / proteinase / フオリプシン
Research Abstract

A 80kDa proteinase, follipsin, was isolated from the follicular fluid of porcine ovaries. The enzyme hydrolyzed internal Arg-X bonds of synthetic and natural peptide substrates, and was highly homologous wiht human plasma kallikrein. The enzyme was strongly inhibited by aprotinin, benzamidine, leupeptin, antipain, and diisopropylfluorophosphate, indicating that it is a serine proteinase. It was found that human single-chain tissue-type plasminogen activator was efficiently converted to its two-chain form by the enzyme. The finding suggests the involvement of the serine proteinase in triggering the activation of proteolytic cascade plasminogen activator/plasmin system, that is generally thought to operate in follicle rupture during ovulation.
Porcine profollipsin was isolated from the fluid. It was readily activated by trypsin treatment. The study also showed that the follicular fluid of porcine ovary contains a profollipsin-activating enzyme activity.
Follicular fluid from human ovaries obtained in in vitro fertilization procedure contained an enzyme similar to porcine follipsin as detected with synthetic, Arg-containing endopeptidase substrates. Purification and characterization studies showed that the enzyme was plasma kallikrein. Plasma kallikrein in association with the proteinase inhibitor alpha 2-macroglobulin was demonstrated to be a predominant form of the enzyme in the fluid, suggesting that the enzyme activity is regulated by the inhibitor. The study also indicated that the plasma kallikrein present in follicular fluid probably comes from the liver.

Report

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

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Kihara,T.: "Identification and activation of profollipsin,a latent precursor of porcine follipsin" Eur.J.Biocchem.(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] Takahashi,T.: "Prolyl endopeptidase from the follicular fluid of porcine ovary: comparison with the liver ezyme" Biomed.Res.17・6. 435-442 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takahashi,T.: "Follipsin,Academic Press" The Handbook of Proteolytic Enzymes (Barrett,A.J.and Woessner,F.Jr.,eds),Academic press. (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] Takahashi,T.: "Proteases involved in follicle rupture during ovulation" J.Reprod.Develop.(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] Ohnishi,J.: "Follicular fluid from human ovaries contains plasma kallikrein in free and in complexed with α 2-macroglobulin" Biomed.Res.,. (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] Kihara, T., Ohnishi, J., Kohyama, K., and Takahashi, T.: "Identification and activation of profollipsin, a latent precursor of porcine follipsin" Eur.J.Biochem.(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] Takahashi, T., Athaua, S.B.P., Mori, T., Kawashima, S., Matsushima, M., Ichinose, M., Miki, K., and Takahashi, K.: "Prolyl endopeptidase from the follicular fluid of porcine ovary : comparison with the liver enzyme" Biomed.Res.17-6. 435-442 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takahashi, T.and Ohnishi, J.: "Follipsin" The Handbook of Proteolytic Enzymes (Barrett, A.J.and Woessner, F., Jr., eds). (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] Takahashi, T., Ohnishi, J., and Kihara, T.: "Proteases involved in follicle rupture during ovulation" J.Reprod.Develop.(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] Ohnishi, J., Murata, M., Kohyama, K., Yoshida, H., Wada, S., Makinoda, S., Fujimoto, S., and Takahashi, T.: "Follicular fluid from human ovaries contains plasma kallikrein in free and in complexed with alpha 2-macroglobulin" Biomed.Res.(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] Kihara, T.: "Identification and activation of profollipsin, a latent precursor of porcine follipsin" Eur. J. Biocchem.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takahashi, T.: "Prolyl endopeptidase from the follicular fluid of porcine ovary : comparison with the liver ezyme" Biomed. Res.17・6. 435-442 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takahashi, T.: "Follipsin, Academic Press" The Handbook of Proteoltic Enzymes (Barrett, A. J. and Woessner, F., Jr., eds)、Academic Press. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takahashi, T.: "Proteases involved in follicle rupture during ovulation" J. Reprod. Develop.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ohnishi,J.: "Cleavage Specificity of Porcine Follipsin" J.Biol.Chem.270. 19391-19394 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takahashi,T.: "Molecular Mechanism of Follicle Rupture during Ovulation" Zool.Sci.12. 359-365 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ohnishi,J.: "Occurrence of a Latent Serine Protease in the Follicular Fluid of Porcine Ovary" Zool.Sci.12. 87-90 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ohnishi,J.: "A Serine Proteinase from the Follicular Fluid of Mammalian Ovaries" Frontiers in Endocrinology. 13. 191-197 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 大西淳之: "排卵機序に関わるプロテアーゼ" 日本比較内分泌学会ニュース. 76. 13-19 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hamabata,T.: "Purification,Characterization,and Localization of Follipsin,ANovel Serine Proteinase from the Fluid of Porcine Ovarian Follicles" J.Biol.Chem.269. 17899-17904 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 大西淳之: "排卵機序に関わるプロテアーゼ" 日本比較内分泌学会ニュース(トピックス). 76. 13-19 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ohnishi,J.: "Occurrence of a Latent Serine Protease in the Follicular Fluid of Porcine Ovary" Zool.Sci.12(印刷中). (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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