| Project/Area Number |
10044198
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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 |
Biophysics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TOYOSHIMA Chikashi Institute of Molecular and Cellular Biosciences, The University of Tokyo, Professor, 分子細胞生物学研究所, 教授 (70172210)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASAKO Masayoshi Institute of Molecular and Cellular Biosciences, The University of Tokyo, Lecturer, 分子細胞生物学研究所, 講師 (30227764)
STOKES David スカボール分子医学研究所, ニューヨーク大学医学センター細胞生物学教室, 準教授
INESI Giusep メリーランド大学, 医学部・生化学分子生物学教室, 教授
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | active transport / protein crystal / membrane proteins / ion pump / over expression / electron crystallography / X-ray crystallography / ATPase |
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| Research Abstract |
The aim of this project is to elucidate the mechanism of active transport from the structural standpoint. When this project was proposed, we had merely two kinds of crystals of CaィイD12+ィエD1-ATPase useful for electron microscopy. Since then, we succeeded in generating large crystals suitable for X-ray crystallography and expected to solve the structure at an atomic resolution within two years or so. Therefore, we shifted the stress on considering on the utilization of mutants for structural studies and asked Prof. Inesi of the University of Maryland to join, because his group was the only one that-succeeded in making large quantity of this membrane protein. This collaboration went out extremely well and produced one review and one original research paper (submitted to Biochemistry). The mutational studies agreed very well with the structure revealed by x-ray crystallography (submitted to Nature) and provided insights into the conformational changes required for active transport. On the other hand, the structure was indispensable to interpret the biochemical and mutational results correctly. Altogether, this project became a very good example of international collaboration. The next step is obviously to make crystals of mutants. For this purpose, the purification procedure used for SR CaィイD12+ィエD1-ATPase has turned out to be insufficient. We are now improving the methods for cultured cells in collaboration with Prof. Inesi.
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