| Project/Area Number |
24659114
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Kyoto University |
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Principal Investigator |
KANEKO Shuji 京都大学, 薬学研究科(研究院), 教授 (60177516)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAGAWA Takayuki 京都大学, 大学院薬学研究科, 准教授 (30303845)
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| Co-Investigator(Renkei-kenkyūsha) |
SHIRAKAWA Hisashi 京都大学, 大学院薬学研究科, 助教 (50402798)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | イオン輸送 / トランスポータ / 細胞内小器官 / シナプス小胞 / 自己組織化単分子層 / 電気生理学 |
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| Research Abstract |
A novel electrophysiological assay technique based on solid supported membranes (SSM) has been shown to be useful for analysis of channels and transporters. We tried to apply this technique to synaptic vesicles (SVs) and lysosomal membranes (LMs), purified from rat brains and mouse livers, respectively. Application of ATP to the sensors with SVs triggered current responses dependent on Mg2+ and sensitive to bafilomycin A1 (Baf), which are thought to be V-ATPase specific currents. On the other hand, though LMs are also embedded with V-ATPases, the sensors evoked current responses with different characteristics. They were independent of Mg2+, only partially sensitive to Baf and even triggered by application of ADP. These features suggest that the currents are not attributed to V-ATPases but any other lysosomal protein. The present study shows the utility and potential of SSM-based electrophysiology especially for analyzing native organellar membrane proteins.
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