| Project/Area Number |
23370047
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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 | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Chiba University |
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Principal Investigator |
MURATA Takeshi 千葉大学, 理学(系)研究科(研究院), 准教授 (80415322)
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| Co-Investigator(Kenkyū-buntansha) |
YAMATO Ichiro 東京理科大学, 基礎工学部, 教授 (70111458)
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| Co-Investigator(Renkei-kenkyūsha) |
SAIJO Shinya 東京理科大学, 基礎工学部, 助教 (80419001)
MIZUTANI Kenji 東京理科大学, 基礎工学部, 博士研究員 (10525570)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥14,950,000 (Direct Cost: ¥11,500,000、Indirect Cost: ¥3,450,000)
Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2011: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
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| Keywords | 分子モーター / 酵素反応 / V-ATPase / X線結晶構造解析 / ATP |
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| Research Abstract |
Vacuolar ATPases (V-ATPases) function as proton pumps in various membrane systems within the cells, which are involved in a number of processes such as bone resorption and cancer metastasis. The hydrophilic V1 portion is known as a rotary motor, in which a central axis DF complex rotates inside a hexagonally arranged catalytic A3B3 complex using ATP hydrolysis energy, although the molecular mechanism is not well defined due to a lack of high-resolution structural information.
We have established the in vitro expression, purification, and reconstitution of Enterococcus hirae V1-ATPase from the A3B3 and DF complexes. In this study, we solved the crystal structures of AMP-PNP-bound and Pi-bound V1-ATPase, and also characterized the several site-directed mutants. On the basis of these findings, we proposed the molecular mechanism of the rotary motor.
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