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An assay method of an endogenous ouabain-like substance

Research Project

Project/Area Number 61870014
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field General pharmacology
Research InstitutionFaculty of Pharmaceutical Sciences, Osaka University

Principal Investigator

IWATA Heitaroh  Faculty of Pharmaceutical Sciences, Osaka University, Prof., 薬学部, 教授 (30028823)

Co-Investigator(Kenkyū-buntansha) 松田 敏夫  大阪大学, 薬学部, 助手 (00107103)
BABA Akemichi  Faculty of pharmaceutical Sciences, Osaka University, Assist, Prof., 薬学部, 教授 (70107100)
MA@TS@UDA Toshio  Faculty of Pharmaceutical Scieneces, Osaka University, Insteructor
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1987: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1986: ¥3,500,000 (Direct Cost: ¥3,500,000)
Keywords(Na^+ + K^+)-ATPase / ouabvain-like factor / phorshorylation / (【Na^+】+【K^+】)-ATPase / ウワバイン様物質 / 内因性物質
Research Abstract

In order to derermine an endogeneous ouabain-like factor, ouabain-stimulated phosphorylation of (Na^+ + K^+)-Atpase by^<32> pi was extensively studied. Sewveral experiments indicate that two isozymes (<alpha>(+) and <alpha>) of (Na^+ + K^+))-Atpase differ in their conformational changes. Furthermore, there was a difference between the two izosymes in the phosphorylation: the Mg^<2+>-dependent phosphorylation by^<32> pi in the absence of ouabain was much less in the <alpha>(+) than in the <alpha>, and sensitivity of the ouqbain-stimulation was much higer in the <alpha>(+) than in the <aplha>. Althousgh the phosphorylation was inhibited by K^+ and cols pi, these ions could be removed from the sample by the pretreatment with a C18-Seppak cartigidge and an ion exchange filter. The, it is concluded that the phosphorylation of the <aplha>(+) is useful for a highly sensitive determination of an endogenous ouabain-like factor. We faild to purify the fdactor from rat plasma using on ODS-HPLC, since the amount of the factor in the plasma was low. Uisng this assay procedure, we determnined the factor level in the plasma of thiamindeficient rate which showed a cardiac dusfunction; the factor level was not changed by thiamin deficiency.

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Toshio Matsuda and Heitaroh Iwata: Arch, Eiochem. Biophys.258. 7-12 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Matsuda and Heitaroh Iwata: Brain Res.437. 375-378 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Heitaroh Iwata; Chizo Iwata and Toshio Matsuda: Japan. J. Pharmacol.46. 35-42 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Matsuda and Heitaroh Iwata: Arch. Biochem. Biophys.(1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Matsuda and %heiraron Iwata: "Phosphorylation of two isozymer of (na^+ + K^+)-ATPase by inorganic phosphate" Arch. Bioche. biophys.258. 7-12 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Matsuda and Heitaroh Iwata: "Decrease of high affinity ouabain binding in rat cerebellum and hyoothalamus by thiamin deficiency" Brain Res.437. 375-378 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Heitaroh Iwata, Chuzo Iwata and Toshio Matsuda: "Differnce between isozymes of (Na^+ + K^+)-ATPase in the interaction with omeprazole" Japan. J. Pharmacol.46. 35-42 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Mtsuda and Heitaroh Iwata: "Differnce between nuwronal and non-nuronal (Na^+ + K^+)-ATPases in their conformational equilibrium" Arch. biochem. Biophys.in press. (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshio Matsud;Heitoroh Iwata: Japan.J.Pharmacol.43. 186P-186P (1987)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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