Crystallization and crystallography of proteins at in vivo salt concentrations
| Project/Area Number |
18K04960
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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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| Review Section |
Basic Section 30010:Crystal engineering-related
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| Research Institution | The University of Tokushima |
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Principal Investigator |
SUZUKI Yoshihisa 徳島大学, 大学院社会産業理工学研究部(理工学域), 准教授 (60325248)
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| Co-Investigator(Kenkyū-buntansha) |
佐藤 正英 金沢大学, 総合メディア基盤センター, 教授 (20306533)
佐崎 元 北海道大学, 低温科学研究所, 教授 (60261509)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | ニワトリ卵白リゾチーム / グルコースイソメラーゼ / 液液相分離 / 平均二乗変位 / 沈殿剤フリー / 生理食塩水 / 分子クラウディング / 平均二乗偏差 / 斜方晶系結晶 / 正方晶系結晶 / 生体内 / タンパク質 / 結晶化 / 結晶構造解析 |
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| Outline of Final Research Achievements |
We have clarified significant structure changes in three-dimensional structure of protein molecules in crystals from different growth conditions. We studied effects of the difference in precipitant condition on the structure, and published papers on the result. In addition, in dilute precipitant conditions, liquid-liquid phase separation frequently occurred during centrifugal concentration of solutions. In the dense liquid phase after the separation, nucleation and growth of crystals occurred very rapidly. Growth rates of obtained crystals in the dense liquid were at least an order of magnitude faster than those in conventional salting-out conditions. In spite of such very rapid processes, maximum resolution limits of obtained crystals were very high (less than 0.1 nm, for instance).
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| Academic Significance and Societal Importance of the Research Achievements |
結晶化条件の違いによって、タンパク質分子の構造が有意に違うということは、創薬等で、生体内の条件下での結晶化が必要とされることを意味する。病因タンパク質の3次元立体分子構造をもとにした特効薬の開発、いわゆるStructrue based drug designでは、そのプロセスで0.15 nmの回折分解能を持つ高品質結晶が要求される。しかし、本研究において、生理食塩水程度の塩濃度における結晶の構造解析、その分解能よりもはるかに大きな構造変化があることが分かった。
本研究では、塩濃度の違いによって明白な構造変化を明らかにし、かつ従来であれば結晶化できないような低塩濃度における結晶化手法を確立した。
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Report
(4 results)
Research Products
(23 results)
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[Journal Article] Highly purified glucose isomerase crystals under microgravity conditions grow as fast as those on the ground do2020
Author(s)
Yoshihisa Suzuki, Ai Ninomiya, Seijiro Fukuyama, Taro Shimaoka, Masae Nagai, Koji Inaka, Shin-ichiro Yanagiya, Takehiko Sone, Shingo Wachi, Yasutomo Arai, Katsuo Tsukamoto
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Journal Title
ChemRxiv.
Volume: preprint
Pages: 1-12
DOI
Related Report
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[Journal Article] Very low nucleation rates of glucose isomerase crystals under microgravity in the International Space Station2019
Author(s)
Yoshihisa Suzuki, Takahisa Fujiwara, Katsuo Tsukamoto, Seijiro Fukuyama, Taro Shimaoka, Kazuma Tsuchiya, Ai Ninomiya, Yumiko Murakumo, Akiho Ikeuchi, Hitomi Minamizono, Tomokazu Yamazaki, Takehiko Sone, Tetsuya Sakashita, Masae Nagai and Yasutomo Arai
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Journal Title
Crystals
Volume: 9
Issue: 2
Pages: 90-90
DOI
NAID
Related Report
Peer Reviewed / Open Access
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[Presentation] Precipitant-free crystallization of proteins2018
Author(s)
Yoshihisa Suzuki, Takahisa Fujiwara, Hideaki Tsuge, Hironori Hondoh, Yusuke Kato, Yuta Uehara, Nobuo Maita, Kohei Hosokawa, Shoko Ueta
Organizer
International Symposium & School on Crystal Growth Fundamentals
Related Report
Int'l Joint Research
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