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Energy-Transducing Enzymes of Higher Plant Vacuolar Membrane

Research Project

Project/Area Number 01560080
Research Category

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

Allocation TypeSingle-year Grants
Research Field 応用生物化学・栄養化学
Research InstitutionHokkaido University

Principal Investigator

MAESHIMA M.  Hokkaido University, Inst. Low Temp. Sci., Associate Professor, 低温科学研究所, 助教授 (80181577)

Co-Investigator(Kenkyū-buntansha) YOSHIDA S.  Hokkaido University, Inst. Low Temp. Sci., Professor, 教授 (90001651)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsProton pump / Pyrophosphatase / Adenosinetriphosphatase / Vacuolar membrane / Magnesium ion / Calcium ion / Subunit / アデノシントリホスファタ-ゼ
Research Abstract

Plant vacuolar membrane contains two distinct proton pumps, namely inorganic pyrophosphatase and ATPase. The vacuolar pyrophosphatase catalyses the PPi hydrolysis and the proton translocation into vacuole. Two proton pumps create a low internal pH and provide an energy source of the secondary transport systems of the vacuolar membrane. In this study, the molecular properties of the enzymes were investigated.
The vacuolar pyrophosphatase was purified from mung bean hypocotyls and shown to consist of a single protein of 73 kDa. An antibody to the 73-kDa protein inhibited the PPi hydrolysis of enzyme. DCCD, which is thought to inhibit ATPases by blocking proton conductance, binds to the 73-kDa protein and inhibits the pyrophosphatase. Thus the 73-kDa protein is an actual H^+-translocating pyrophosphatase. The pyrophosphatase functions as a homodimer comprised of 73-kDa subunit. The enzyme requires Mg^<2+> for its stability and activity. Ca^<2+>, however, strongly inhibits the pyrophosphatase. The rate of inhibition by Ca^<2+> depended on the concentration of Ca-PPi complex, and the apparent K_i for CaPPi was about 17 uM.
The vacuolar ATPase was also purified from mung bean and shown to consist of nine different subunits (68, 57, 44, 38, 37, 32, 16, 13 and 12 kDa). Except for the 16-kDa subunit, the other subunits were released from the membrane by treatment with KSCN. These eight subunits may compose the hydrophilic part of the ATPase, and the DCCD-binding subunit of 16 kDa composes the proton channel. Enzymatic properties of the ATPase and pyrophosphatase were also analyzed.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Maeshima,M.: "Purification and properties of vacuolar membrane protontranslocating iorganic pyrophpsphatase from mung bean." Journal of Biological Chemistry. 264. 20068-20073 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] MatsuuraーEndo,C.: "Subunit composition of vacuolar membrane H^+ーATPase from mung bean." European Journal of Biochemistry. 187. 745-751 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Maeshima,M.: "Development of vacuolar membranes during elongation of cells in mung bean hypocotyls." Plant & Cell Physiology. 31. 311-317 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Maeshima,M.: "Oligomeric structure of H^+ーtranslocating inorgnic pyrophosphatase of plant vacuoles." Biochemical and Biophysical Research Communications. 168. 1157-1162 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Maeshima,M.: "H^+ーtranslocating inorganic pyrophosphatase of plant vacuoles: Inhibition by Ca^<2+>,stabillzation by Mg^<2+> and immunological comparison with other inorganic pyophsophatases." European Journal of Biochemistry. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 前島 正義: "植物細胞の活動を支える液胞膜の機能" 植物細胞工学. 2. 79-83 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Maeshima,M. (分担執筆): "Plant Water Relations and Growth under Stress" Myu Co.,Tokyo, (507)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Masayoshi Maeshima, & S. Yoshida: "Purification and properties of vacuolar membrane proton-translocating inorganic pyrophosphatase from mung bean." J. Biol. Chem.264. 20068-20073 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Matsuura-Endo, C., Masayoshi Maeshima, & S. Yoshida: "Subunit composition of vacuolar membrane HATPase from mung bean" Eur. J. Biochem.187. 745-751 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Masayoshi Maeshima: "Development of vacuolar membrane during elongation on cells in mung bean hypocotyls." Plant & Cell Physiol.31. 311-317 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Masayoshi Maeshima: "Oligomeric structure of H^+-translocating inorganic pyrophosphatase of plant vacuoles." Biochem. Biophys. Res. Commun.168. 1157-1162 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Masayoshi Maeshima: "H^+-translocating inorganic pyrophosphatase of plant vacuoles : Inhibition by Ca^<2+>, stabilization by Mg^<2+> and immunological comprison with other inorgnic pyrophosphatases." Eur. J. Biochem.196. 11-17 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Maeshima,M.: "Purification and properties of vacuolar membrane protonーtranslocating iorganic pyrophosphatase from mung bean." Journal of Biological Chemistry. 264. 20068-20073 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] MatsuuraーEndo,C.: "Subunit composition of vacuolar membrane H^+ーATPase from mung bean." European Journal of Biochemistry. 187. 745-751 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeshima,M.: "Development of vacuolar membranes during elongation of cells in mung bean hypocotyls." Plant & Cell Physiology. 31. 311-317 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeshima,M.: "Oligomeric structure of H^+ーtranslocating inorgnic pyrophosphatase of plant vacuoles." Biochemical and Biophysical Research Communications. 168. 1157-1162 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeshima,M.: "H^+ーtranslocating inorganic pyrophosphatase of plant vacuoles:Inhibition by Ca^<2+>,stabilization by Mg^<2+> and immunological comparison with other inorganic pyophsophatases." European Journal of Biochemistry. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 前島 正義: "植物細胞の活動を支える液胞膜の機能" 植物細胞工学. 2. 79-83 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Maeshima,M.(分担執筆): "Plsnt Water Relations and Growth under Stress" Myu Co.,Tokyo, 507 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] MAESHIMA Masayoshi: "Purification and properties of vacuolar membrane protontranslocationg inorganic pyrophosphatase from mung bean" Journal of Biological Chemistry. 264. 20068-20073 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] MATUUTA-ENDO Chie: "Subunit composition of vacuolar membrane H^+-ATPase from mung bean" European Journal of Biochemistry. 187. 745-751 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] MAESHIMA Masayoshi: "Development of vacuolar membranes during elongation of cells in mung bean hypocotyls" Plant & Cell Physiology. 31. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] MAESHIMA Masayoshi: "Inhibition of proton-translocating inorganic pyrophosphatase of vacuolar membrane by calcium" European Journal of Biochemistry.

    • Related Report
      1989 Annual Research Report
  • [Publications] 前島正義: "植物細胞の活動をささえる液胞膜の機能" 植物細胞工学. 2. 79-83 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] MAESHIMA Masayoshi: "Plant Water Relations and Growth under Stress(Proceedings of the Yamada Conference XXII)分担執筆(pp.407-409)" Myu K.K.(Japan), 507 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 前島正義: "現代植物生理学,第2巻(全5巻)“エネルギ-代謝"(分担執筆)" 朝倉書店,

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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