An assay method of an endogenous ouabain-like substance
| Project/Area Number |
61870014
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Faculty of Pharmaceutical Sciences, Osaka University |
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Principal Investigator |
IWATA Heitaroh Faculty of Pharmaceutical Sciences, Osaka University, Prof., 薬学部, 教授 (30028823)
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| 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
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| 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)
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| Keywords | (Na^+ + K^+)-ATPase / ouabvain-like factor / phorshorylation / (【Na^+】+【K^+】)-ATPase / ウワバイン様物質 / 内因性物質 |
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| 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.
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Report
(2 results)
Research Products
(9 results)
