| Project/Area Number |
26893117
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Physical pharmacy
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| Research Institution | Kyoto University |
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Principal Investigator |
Kawano Kenichi 京都大学, 物質-細胞統合システム拠点, 研究員 (70732874)
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| Project Period (FY) |
2014-08-29 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 可溶化剤 / M2プロトンチャネル / S31N変異体 / 溶液NMR / A型インフルエンザウイルス / アマンタジン耐性 / 二量体 |
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| Outline of Final Research Achievements |
The synthesis and purification methods of a novel detergent were established. Before applying it to M2 protein, the well-studied photosensitive bacteriorhodopsin (bR) was used as a model membrane protein to examine the solubilization ability of the detergent, and the time-dependent thermal stability of bR in the detergent micelle, monitoring change in the absorbance spectrum. As a result, the abilities of the detergent were proven to be significantly improved compared with the existing detergents. The micelle size after solubilizing bR was available for solution NMR. Although the expression of M2 protein was confirmed by the Western blotting, only a part of M2 protein formed the dimers crosslinked by the disulfide bonds at the N-termini. To facilitate the crosslinking, a self-associating coiled-coil sequence was inserted into the N-terminus of the M2 protein. The structural analysis of the dimeric M2 protein will be started following confirmation of M2 dimerization.
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