2023 Fiscal Year Final Research Report
Development of fixed target micro-crystallography dedicated to structural dynamics study
| Project Area | Non-equilibrium-state molecular movies and their applications |
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| Project/Area Number |
19H05783
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| Research Category |
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
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| Allocation Type | Single-year Grants |
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| Review Section |
Complex systems
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Yamamoto Masaki 国立研究開発法人理化学研究所, 放射光科学研究センター, 部門長 (60241254)
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| Co-Investigator(Kenkyū-buntansha) |
熊坂 崇 公益財団法人高輝度光科学研究センター, 構造生物学推進室, 室長 (30291066)
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| Project Period (FY) |
2019-06-28 – 2024-03-31
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| Keywords | X線結晶構造解析 / 構造ダイナミクス / 放射光 |
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| Outline of Final Research Achievements |
We have developed a vacuum diffractometer for high-precision data collection on a fixed target and an in-situ measurement system for the generalization of dynamic X-ray crystallography. The vacuum diffractometer was found to be effective for low-background high S/N measurements, as the background was reduced by 99.9% under vacuum conditions compared to atmospheric conditions. The high-temperature humidity controller for the in-situ measurement system was used to control humidity up to 70°C and detect changes in protein structure and hydration structure. In addition, an injector-based microcrystal room temperature serial data collection combining the rtHAG and SS-ROX methods was successfully used for structural analysis. We also succeeded in developing a crystallization method suitable for humidity conditioning measurements to create different crystal systems depending on the scaffold protein.
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| Free Research Field |
構造生物学
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| Academic Significance and Societal Importance of the Research Achievements |
構造ダイナミクス研究に向けXFELビームタイムの確保が律速となるSFX法に加え、リング型放射光でも固定した多量の微小結晶のからの動的Ｘ線結晶構造解析の高度化が求められている。真空回折計は低バックグラウンド高S/N測定を実現して、構造変化の同期に有利なナノメートルサイズ結晶からの動的Ｘ線結晶構造解析を可能にする。また、非凍結での構造解析を実現するin-situ測定系は、恒温動物の体温域まで測定範囲を拡張しSS-ROX法と組み合わせることでタンパク質の生理活性温度環境での構造決定を可能にした。これらの技術開発により、広範な測定条件でのタンパク質の動的Ｘ線結晶構造解析を可能にする。
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