| Project/Area Number |
25440037
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | National Institutes for Quantum and Radiological Science and Technology (2016)
Japan Atomic Energy Agency (2013-2015) |
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Principal Investigator |
Tamada Taro 国立研究開発法人量子科学技術研究開発機構, 高崎量子応用研究所 東海量子ビーム応用研究センター, 上席研究員(定常) (50391248)
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| Co-Investigator(Kenkyū-buntansha) |
平野 優 国立研究開発法人量子科学技術研究開発機構, 高崎量子応用研究所 東海量子ビーム応用研究センター, 研究員(定常) (80710772)
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| Co-Investigator(Renkei-kenkyūsha) |
KIMURA Shigenobu 茨城大学, 工学部, 教授 (90291608)
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| Project Period (FY) |
2013-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 酸化還元酵素 / 高分解能Ｘ線結晶構造解析 / 中性子結晶構造解析 / 高分解能X線結晶構造解析 / 時分割Ｘ線結晶解析 |
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| Outline of Final Research Achievements |
NADH-cytochrome b5 reductase (b5R), a flavoprotein consisting of NADH- and FAD- domains, catalyzes electron transfer from the two-electron carrier NADH to the one-electron carrier cytochrome b5 (b5). We succeeded in structure determination of b5R and b5 at several states during the catalytic cycle by high-resolution X-ray crystallography. In the b5R oxidized (OX) form, valence electrons were clearly visualized and analyzed by multipolar refinement at 0.78Å resolution. In the b5R reduced (RD) form, the relative configuration of the two domains was shifted in comparison with the OX form, resulting in the creation of a new hydrogen bonding network around FAD. Time-resolved analysis of b5R suggested that re-oxidation follows a two-step mechanism. We also confirmed the slight structural change, which regulates the catalytic cycle, between the OX and the RD form of b5 above 1.0Å resolution. In addition, we collected the neutron diffraction data of the OX form of b5R at 1.4Å resolution.
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