Protein 19F Labeling Strategy for In-Cell NMR and Functional Analyses
| Project/Area Number |
26440018
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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 | Tohoku University |
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Principal Investigator |
Koshiba Seizo 東北大学, 東北メディカル・メガバンク機構, 教授 (70332301)
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| Project Period (FY) |
2014-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | フッ素 / タンパク質 / NMR / 細胞内環境 / 相互作用 / 動的構造変化 / 19F-NMR / In-Cell NMR / 動的構造 |
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| Outline of Final Research Achievements |
Fluorine 19 (19F) is a widely used nucleus for NMR because of its high sensitivity and large chemical shift dispersion. In particular, 19F-labeling is useful for In-Cell NMR, because most of biological systems contain no fluorine, resulting that 19F-labeled molecules in living cells can be selectively analyzed. In this study, I developed several 19F-labeling methods for proteins and effectively incorporated trifluoromethyl-labeled amino acids into proteins. I expressed several kinds of trifluoromethyl-labeled proteins and could observe highly sensitive 19F NMR signals both in vitro and in vivo. I also investigated the interactions of ligands with large molecular weight proteins by 19F-NMR and could identify binding sites of the ligands in a short time. These results show that my methods are particularly useful for investigating behavior of intracellular proteins or ligand-interactions. I expect that this labeling strategy is useful for wide variety of biological sciences.
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Report
(5 results)
Research Products
(17 results)
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[Journal Article] Evaluation of reported pathogenic variants and their frequencies in a Japanese population based on a whole-genome reference panel of 2,049 individuals2018
Author(s)
Y Yamaguchi-Kabata, J Yasuda, O Tanabe, Y Suzuki, H Kawame, N Fuse, M Nagasaki, Y Kawai, K Kojima, F Katsuoka, S Saito, I Danjoh, I Motoike, R Yamashita, S Koshiba, D Saigusa, G Tamiya, S Kure, N Yaegashi, Y Kawaguchi, F Nagami, S Kuriyama, J Sugawara, N Minegishi, A Hozawa, T Takai-Igarashi, K Kinoshita, M Yamamoto.
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Journal Title
Journal of Human Genetics.
Volume: 63
Issue: 2
Pages: 213-230
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] The structural origin of metabolic quantitative diversity.2016
Author(s)
Koshiba S, Motoike I, Kojima K, Hasegawa T, Shirota M, Saito T, Saigusa D, Danjoh I, Katsuoka F, Ogishima S, ... Yamamoto M.
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Journal Title
Sci Rep
Volume: 6
Issue: 1
Pages: 31463-31463
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Establishment of Protocols for Global Metabolomics by LC-MS for Biomarker Discovery.2016
Author(s)
Saigusa D, Okamura Y, Motoike IN, Katoh Y, Kurosawa Y, Saijyo R, Koshiba S, Yasuda J, Motohashi H, Sugawara J, Tanabe O, Kinoshita K, Yamamoto M
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Journal Title
PLoS One
Volume: 11
Issue: 8
Pages: e0160555-e0160555
DOI
Related Report
Peer Reviewed / Open Access
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