| Project/Area Number |
16K15179
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
General physiology
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| Research Institution | University of Fukui |
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Principal Investigator |
Oiki Shigetoshi 福井大学, 学術研究院医学系部門, 教授 (10185176)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | イオンチャネル / 脂質２重膜 / リン脂質 / コレステロール / in vitro転写/翻訳 / 脂質二重膜 / 膜張力 / in vitro転写・翻訳系 / 脂質平面膜 / カリウムチャネル / イオン透過 / 蛋白質 / 薬理学 / 生理学 |
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| Outline of Final Research Achievements |
To study channel-lipid interplay in the lipid bilayer membrane, we developed several basic methods using the contact bubble bilayer (CBB) method. For the CBB, in vitro transcription/translation system was used, and the KcsA channel proteins were inserted, folded and assembled as functional channels detected as single channel currents. This system provides a versatile platform for channel protein expression-functional studies including the TREK-1 channel. CBB allowed formation of lipid bilayer with arbitrary lipid composition, and we developed the rapid perfusion method. The KcsA potassium channel closed its activation gate upon cholesterol perfusion, and stopping the perfusion led recovery of the channel activity because of dilution of cholesterol in the bilayer. Repeated on and off of the perfusion produced repeated activation and deactivation of the channel gating. This system allows examinations of hydrophobic drug actions to channel proteins.
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