Quantitative assessment of activity of light-driven proton pump bacteriorhodopsin
| Project/Area Number |
61550698
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
化学工学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TODA Kiyoshi Institute of Applied Microbiology, The University of Tokyo, Proffessor, 応用微生物研究所, 教授 (00015637)
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| Co-Investigator(Kenkyū-buntansha) |
YABE Isamu Institute fo Applied Microbiology, The university of Tokyo, Assistant Pro., 応用微生物研究所, 助手 (90111575)
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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 |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1987: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1986: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Pyranine / Fluorescenceanisotropy / Bacteriorhodopsin / Proton pumping rate / Simultaneous pH measurement / 平面リン脂質二分子膜 / パッチ電流法 / スフェロプラスト |
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| Research Abstract |
1)The accuracy of measurement of pH with a pH-sensitive fluorescent probe of pyranine was investigated extensively. The determination of fluorescence anisotropies of the liposomes comprising pyranine indicated that the fraction of pyranine adsorbed on the internal surface of liposomes was several percent of the intravesicular concentration of pyranine. Secondary,it was confirmed that interference by phospholipid in fluorescence measurement of intravesicular pyranine could be compensated simply by subdracting by the background fluorescence. we could determine the intravesicular pH within a standard deviation of 0.01 pH.
2)Simultaneous changes of pH in the inner and outer liquids of proteoliposomes,the membrane of which consisted of asolectin and bacteriorhodopsin(BR),were determined during transient states after a schift from the light to the dark and the reverse. Some mathematical equations of a model characterizing the mechanism of the proton pump of BR, light-dependent and light-independent proton leaks from the liposomes ,and buffering capacities of the intra- and extra-vesicular liquids. The model could simulate well the dynamic aspect of pH changes.
3)The equipment for the measurement of the proton pump current ,which is generated by BR in planar bilayer phospholipid membrane, was developed.
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Report
(2 results)
Research Products
(6 results)
