2013 Fiscal Year Final Research Report
Structural study of H+-translocating inorganic pyrophosphatase
| Project/Area Number |
24770091
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2014-03-31
|
| Keywords | 膜タンパク質 / プロトンポンプ / 高等植物 / 結晶化 / X線結晶解析 |
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| Research Abstract |
Proton-translocating inorganic pyrophosphatase is a membrane protein that uses the energy from the hydrolysis of inorganic pyrophosphate to transport protons across biological membranes. This study used X-ray crystallography to determine the structure of this enzyme without bound
pyrophosphate. The enzyme was purified from the plant vacuolar membrane and crystallized in the absence of pyrophosphate. Crystals were grown in a complex with an antibody fragment that specifically binds to the enzyme in the absence of bound pyrophosphate. The initial crystals showed a diffraction resolution of 4.1 angstrom. This resolution limit was improved to 3.2 angstrom by optimizing the conditions of
crystallization and cryoprotection.
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