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Molecular genetic study on the metabolism and function of phosphatidylserine in mammalian cells

Research Project

Project/Area Number 03671077
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionNational Institute of Health

Principal Investigator

NISHIJIMA Masahiro  NIH, Head, 細胞化学部, 室長 (60072956)

Co-Investigator(Kenkyū-buntansha) SAITO Kyoko  NIH,Researcher, 細胞化学部, 研究員 (70235034)
AKAMATSU Yuzuru  NIH, Director, 細胞化学部, 部長 (00072900)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsPhosphatidylserine / Phosphatidylserine synthase / Phosphatidylserine decarboxylase / Gene targeting / ホスファチジル脱炭酸酵素 / 遺伝子タ-ゲッティング
Research Abstract

To reveal the mechanism of phosphatidylserine (PS) biosynthesis and function of PS, we previously isolated a mammalian cell mutant defective in PS biosyntheses. By using this mutant, we found that PS synthase and PS decarboxylase played important roles in PS formation in mammalian cells. In the present study, to further characterize the molecular mechanism of PS synthesis, we tried to purify PS synthase and to isolate mutants defective in PS decarboxylase.
(1) We made polyclonal antibody to PS synthase by using a oligopeptide which corresponded to the C-terminal part of PS synthase.
(2) We tried to solubilize PS synthase by using various kinds of detergents. However, we could not find good conditions to solubilize the enzyme. (3) We desrupted the gene of PS decarboxylase in CHO cells by gene targeting. In this way, we obtained a mutant defective in one of the Ps decarboxylase genes.
Our results are still premature and so we will continue the above studies for coming several years.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] K.HANADA: "Sphingolipids are essential for the growth of Chinese hamster ovary cells" J.Biol.Chem.267. 23527-23533 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 赤松 穣編集: "脂質の分析法" 廣川書店, 180 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 日本化学会編: "脂質の化学と生化学" 学会出版センター, 177 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kentaro Hanada, Masahiro Nishijima, and Yuzuru Akamatsu: "A temperature-sensitive mammalian cell mutant with thermolabile serine palmitoyltransferace for the sphingolipid biosynthesis" J.Biol. Chem.vol.265. (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Osamu Kuge, Masahiro Nishijima, Yuzuru Akamatsu: "A cloned gene encoding phosphatidylserine decarboxylase complements the phosphatidylserine biosynthetic defect of a Chinese hamster ovary cell mutant" J. Biol. Chem.vol.266. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Osamu Kuge, Masahiro Nishijima, Yuzuru Akamatsu: "A Chinese hamster cDNA encoding a protein essential for phosphatidylserine synthase I activity" J. Biol. Chem.vol.266. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kentaro Hanada, Masahiro Nishijima, Makoto Kiso, Akira Hasegawa, Shuji Fujita, Tomoya,Ogawa, and Yuzuru Akamatsu: "Spingolipids are essential for the growth of Chinese hamster ovary cells" J. Biol. Chem.vol.267. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Osamu kuge, Kiyo Izawa, Masahiro Nishijima, Yuzuru Akamatsu: "Shingolipid deficiency induces hypersensitivity of CD14, a glycosyl phosphatidylinositol-anchored protein, to phosphatidylinositol specific phospholipse C" J. Biol. Chem.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Hanada: "Sphingolipids are essential for the growth of Chinese hamster ovary cells" J.Biol.Chem.267. 23527-23533 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 赤松 穣 編集: "脂質の分析法" 廣川書店, 180 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 日本化学会編: "脂質の化学と生化学" 学会出版センター, 177 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Hanada,M.Nishijima,Y.Akamatsu.: "A temperature-sensitire mamn alian cell mutant with thermolalile sevine palmitoyltransfer rse for the sphin go lipid lios prtheis" J.Biol.Chem.265. 22137-22142 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] O.Kenge,M.Nishijima,Y.Akamatsu: "A cloned gene encoding phosphatidylieinl decavloxylase complements the phosphatidylserine biosyntheti defect of a Chinesl hanstel overy cell matant" J.Biol.Chem.266. 6370-6776 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] O.Kuge,M.Nishijima,Y.Akamatsu: "A Chinese habster cDNA encoding a protein essential for phasphatidylserine synthase I activity" J.Biol.Chem.266. 24184-24189 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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