| Project/Area Number |
17K07317
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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 | Osaka Medical College |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
林 秀行 大阪医科大学, 医学部, 教授 (00183913)
馬場 清喜 公益財団法人高輝度光科学研究センター, タンパク質結晶解析推進室, 主幹研究員 (00437344)
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| Project Period (FY) |
2017-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | トパキノン / HAG法 / 銅含有アミン酸化酵素 / アミン酸化酵素 / X線結晶構造解析 / 蛋白質 / 酵素 / 結晶 / 反応中間体 |
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| Outline of Final Research Achievements |
Many of the protein X-ray crystallographic analyses are currently performed at cryogenic temperatures to reduce the radiation damage. However, the obtained structures are ambiguous in terms of temperature information. In this study, we applied the humid air and glue-coating (HAG) method, which was initially developed as a protein crystal-mounting method applicable to noncryogenic X-ray crystallography, to thermodynamically analyze the conformational change of the cofactor in noncryocooled crystals of bacterial copper amine oxidase. The temperature-controlled HAG method provides a technique for thermodynamic analysis of conforma- tional changes occurring in protein crystals that are hardly scrutinized by conventional cryogenic X-ray crystallography.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究成果により，機能している状態のタンパク質の構造変化を，熱力学的に理解する一般的な手法が開発された．今後さらに，各種タンパク質の機能を，立体構造や平衡状態の変化をもとに解明し，有用酵素の作製や酵素を阻害する薬剤の開発などにも役立たせる基礎的な方法論として発展が期待できる．また，今まで結晶構造は溶液構造と異なり非生理的なのではないかという懸念があったが，今回の解析で，むしろ結晶構造の方が細胞内のタンパク質の状態を反映しているという重要な知見も得られた．
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