| Project/Area Number |
07558220
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Functional biochemistry
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
TANIGUCHI Kazuya HOKKAIDO UNIV., GRADUATE SCHOOL OF SCIENCE,PROFESSOR, 大学院・理学研究科, 教授 (40028204)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRASAKI Yuji MUTOU TECHNOS,CHIEF, 部長
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Na, K-ATPase / Rb.Occlusion / Phosphorylation / キナーゼ / 胃壁H^+ポンプ / ATPase / 脱リン酸化 / カルシウムイオン / Na^+ポンプ / P型ポンプ / ラピッドフィルトレーション / 構造変化 / イオン閉塞 / 蛍光プローブ |
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| Research Abstract |
Pig Kidney Na, K-ATPase preparations modified with N- [p- (2-benzimidazolyl) phenyl] maleimide (BIPM-enzyme) were incubated with 0.7muM N- (4-sulfobutyl) -4- [4- (p-dipentyl-aminophenyl) butadienyl] pyridinium, inner salt (RH421). The resulting enzyme preparations changed each relative fluorescence intensity under conditions of accumulation of various reaction intermediates.
Addition of ATP to the enzyme preparations in the presence of 2M Na^+ and 4 mM Mg^<2+> induced a rapid decrease in RH421 fluorescence (25/s) follwed by slow phosphoenzyme formation and decrease in BIPM fluorescence (5-6/s). The addition of ATP to the preparation in the presence of 16 mM Na^+ and 1 mM Ca^<2+> induced sequential decreases in fluorescence of RH421 (44/s) and BIPM(26/s), which were followed by slow increases in both BIPM fluorescence and phosphoenzyme (around 6/s) and slower increase in RH421 fluorescence (1.7/s). RH421 seemed to sense ATP dependent molecular events preceding BIPM fluorescence decrease prior ADP sensitive phosphoenzyme formation and following K^+ -sensitive phosphoenzyme formation.
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