Establishment of the technical basis for a new "protein cage structure analysis methods" using ferritin
| Project/Area Number |
18K05314
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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 37010:Bio-related chemistry
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
Kanamaru Shuji 東京工業大学, 生命理工学院, 助教 (50376951)
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| Project Period (FY) |
2018-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 単粒子解析 / ケージ蛋白質 / 立体構造解析 / 構造解析 / 蛋白質ケージ / Scaffold / クライオ電顕 / scaｆfold / scafold |
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| Outline of Final Research Achievements |
To determine the structure of proteins smaller than 50 kDa using cryo-EM single-particle analysis, we first constructed an over-expression system in Escherichia coli and succeeded in confining the target protein in the inner cavity of a 24-mer ferritin protein cage. In addition, we succeeded in selectively purifying only the cages in which the target protein was introduced by using appropriate purification tags.
On the other hand, the number of target proteins in the inner cavity could be controlled to some extent by narrowing the inner space, but it was difficult to align the position and orientation of the target protein even by changing the type and length of the linker, and we still could not obtain the high-resolution structure of the target proteins.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究で開発したフェリチン24量体内部空洞に効率的に標的蛋白質を導入し、単離精製する方法は、内部空洞の大きさよりも小さい蛋白質であれば標的蛋白質の配列を変えることなく簡便に導入することが出来る。フェリチンのもつ高い熱安定性・pH安定性を利用することで内部空洞に導入した蛋白質に機能を改変せずに安定性を付加することができると期待される。
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Report
(6 results)
Research Products
(15 results)
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[Journal Article] Cooperative interactions facilitate stimulation of Rad51 by the Swi5-Sfr1 auxiliary factor complex.2020
Author(s)
Argunhan B, Sakakura M, Afshar N, Kurihara M, Ito K, Maki T, Kanamaru S, Murayama Y, Tsubouchi H, Takahashi M, Takahashi H, Iwasaki H.
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Journal Title
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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