| Project/Area Number |
15H03841
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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 |
Bio-related chemistry
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Kubo Minoru 国立研究開発法人理化学研究所, 放射光科学総合研究センター, 専任研究員 (90392878)
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| Co-Investigator(Renkei-kenkyūsha) |
TOSHA Takehiko 国立研究開発法人理化学研究所, 放射光科学総合研究センター, 専任研究員 (00548993)
KIMURA Tetsunari 神戸大学, 大学院理学研究科, 特命講師 (70506906)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2015: ¥13,260,000 (Direct Cost: ¥10,200,000、Indirect Cost: ¥3,060,000)
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| Keywords | 時間分解赤外分光 / マイクロ流路フローセル / 酵素反応 / 膜タンパク質 / 一酸化窒素還元酵素 / 超精密計測 / フェムト秒レーザー / ケージド化合物 / ポンププローブ法 / 蛋白質 / 高性能レーザー / 生物物理 |
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| Outline of Final Research Achievements |
This project aims to establish a time-resolved IR spectroscopic technique for studying dynamic structures of enzymes. A novel micro flow system has been developed and incorporated into a femtosecond laser-based IR spectrometer, which allows micro flow-flash IR measurements of enzymatic reactions with the minimum sample volume of 20 nL. We applied this measurement system to NO reductase, a heme enzyme that catalyzes the reduction of NO to N2O in the nitrogen cycle, and succeeded in determining the NO binding mode in the short-lived intermediate. Furthermore, we also used the micro-flow flash method for time-resolved crystallography at SACLA, and succeeded in visualizing the 3D structure of the NO-bound intermediate during the NO reduction reaction.
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