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STIMULATION AND DISORDER OF PHOSPHATIDYLCHOLINE METABOLISM CELL GROWTH

Research Project

Project/Area Number 05454164
Research Category

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

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionGunma University

Principal Investigator

YAMASHITA Satoshi  Gunma University, Department of Biochemistry, Professor, 医学部, 教授 (50025623)

Co-Investigator(Kenkyū-buntansha) UCHIDA Tsutomu  Gunma University, Department of Biochemistry, Assistant, 医学部, 助手 (00160276)
HOSAKA Kohei  Gunma University, Department of Biochemistry, Associate Professor, 医学部, 助教授 (70108992)
SUGIMOTO Hiroyuki  Gunma University, Department of Biochemistry, Assistant, 医学部, 助手 (00235897)
OKAMURA Shinichi  Gunma University, Department of Biochemisty, Assistant, 医学部, 助手 (20224058)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1994: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1993: ¥5,200,000 (Direct Cost: ¥5,200,000)
Keywordsphosphatidylcholine metabolism / cell growth / signal transduction / enzyme purification / phospholipase D / lysophospholipase / phosphatidic acid / lysophosphatidylcholine / リゾホスファチジルコン / コリンキナーゼ
Research Abstract

Phosphatidylcholine is profoundly involved in the signal transduction and growth of cells. We are interested in the roles of phosphatidylcholine metabolism in cell proliferation. The presen study dealt with the purification, and cloning of cDNA of phospholipases in hosphatidylcholine metabolism. The major results can be summarized as follows.
Phospholipase D,believed to play a central role in the cell growth signaling pathway, was purified 2,200-fold to homogeneity from pig liver microsomes by a combination of column chromatographies. The molecular weight was determined to be 190,000 by SDS electrophoresis. Lysophospholipases acting on lysophosphatidylcholine were purified to homogeneity from rat liver and pig gastric mucosa. From liver, two isoforms with molecular masses of 24 and 60 kd were obtained. The latter enzyme exhibited not only hydrolase activity, but also transacylase activity. From gastric mucosa, two lysophospholipases were also obtained with molecular masses of 22 and 23 kd. Antibodies were raised against hepatic 24-kd and gastric 22- and 23-kd enzymes. Western blot analysis showed that the liver 24-kd enzyme was immunologically related to gastric 22-kd enzyme, but not to liver 60-kd and gastric 23-kd lysophospholipases. All these lysophospholipases were inhibited by phosphatidic acid and lysophosphatidic acid to varying extents. We obtained cDNA clone for hepatic 24-kd lysophospholipase from lambda gtl 1 liver cDNA library using the antibody. The cDNA clone contained the nucleotide sequence corresponding to the partial amino acid sequences determined for the purified hepatic 24-kd lysophospholipase.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report

Research Products

(21 results)

All Other

All Publications (21 results)

  • [Publications] J.Nikawa: "Differential expression of two myo-inositol transporter genes of Saccharomyces cerevisiae" Mol.Microbiol.10. 955-961 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Hosaka: "Cloning and characterization of the SCS1 gene rrequired for the expression of genes in yeast phospholipid synthesis" J.Biochem.115. 131-136 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] H.sugimoto: "Purification,characterication,and inhibition by phosphatidic acid of lysophosppholipase-transacylase from rat liver" J.Biol.Chem.269. 6252-6258 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Hosaka: "Cloning and sequence of the SCS3 gene which is required for inositol prototrophy in Saccharomyces cerevisiae" J.Biochem.116. 1317-1321 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Okamura: "Purification and characterication of phosphatidylcholine phospholipase D from pig lung" J.Biol.Chem.269. 31207-31213 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kodaki: "The SNF2/SWI2/GAM1/TYE3/RIC1 gene is involved coordinate regulation of phospholipid synthesis in Saccharomyces cerevisia" J.Biochem.1995 (117)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kodaki: "The SNF2/SWI2/GAM1/TYE3/RIC1 gene is involved coordinate regulation of phospholipid synthesis in Saccharomyces cerevisia" J.Biochem.1995 (117)362-368:

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] J.Nikawa, K.Hosaka, and S.Yamashita: "Differential expression of two myo-inositol transporter genes of Saccharomyces cerevisiae" Mol.Microbiol.10. 955-961 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Hosaka, J.Nikawa, T.Kodaki, and S.Yamashita: "Cloning and characterization of the SCS1 gene required for the expression of genes in yeast phospholipid synthesis" J.Biochem.115. 131-136 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] H.Sugimoto and S.Yamashita: "Purification, characterization, and inhibition by phosphatidic acid of lysophospholipase-transacylase from rat liver" J.Biol.Chem.269. 6252-6258 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Hosaka, J.Nikawa, T.Kodaki, and S.Yamashita: "Cloning and sequence of the SCS3 gene which is required for inositol prototrophy in Saccharomyces cerevisiae" J.Biochem.116. 1317-1321 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Okamura and S.Yamashita: "Purification and characterization of phosphatidylcholine phospholipase D from pig lung" J.Biol.Chem. 269. 31207-31213 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Kodaki, K.Hosaka, J.Nikawa, and S.Yamashita: "The SNF2/SW12/GAM1/TYE3/RIC1 gene is involved in the coordinate regulation of phospholipid synthesis in Saccharomyces cereviside" J.Biochem.117. 362-368 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] H.Sunaga, H.Sugimoto, Y.Nagamachi, and S.Yamashita: "Purification and properties of lysophospholipase isozymes from pig gastric mucosa" Biochem.J.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Okamura: "Purification and characterization of phosphatidylcholine phospholipase D from pig lung" J. Biol. Chem.269. 31207-31213 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Hosaka: "Cloning and sequence of the SCS3 gene which is required for inositol prototrophy in Saccharomyces cerevisiae" J. Biochem. 116. 1317-1321 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] H.Sugimoto: "Purification, characterization, and inhibition by phosphatidic acid of lysophospholipase-transacylase from rat liver" J. Biol. Chem.269. 6252-6258 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Kodaki: "The SNF2/SWI2/GAM1/TYE3/RIC1 gene is involved in the coordinate regulation of phospholipid synthesis in Saccharomyces cerevisia" J. Biol.Chem. 117. 362-368 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] J.Nikawa: "Differential expression of two myoinositol transporter genes of Saccharomyces cerevisiae" Mol.Microbiol.10. 955-961 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Hosaka: "Cloning and characterization of the SCS1 gene required for the expression of genes in yeast phospholipid synthesis" J.Biochem.115. 131-136 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Sugimoto: "Purification,characterization,and inhibition by phosphatidic acid of lysophospholipase-transacylase from rat liver" J.Biol.Chem.269(印刷中). (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-03-31   Modified: 2016-04-21  

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