| Project/Area Number |
16K15105
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Physical pharmacy
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| Research Institution | Kyoto University |
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Principal Investigator |
Kato Hiroaki 京都大学, 薬学研究科, 教授 (90204487)
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| Co-Investigator(Renkei-kenkyūsha) |
MIYANOIRI Yohei 大阪大学, 蛋白質研究所, 准教授 (80547521)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 膜タンパク質 / NMR / 多剤耐性 / ATP / トランスポーター / 構造生物学 / 薬理学 / 好熱性真核生物 |
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| Outline of Final Research Achievements |
P-glycoprotein is an ATP Binding Cassette multidrug transporter which is a large membrane protein that works in eukaryotic cells. Thus, it is difficult to have a large amount of its 15N-labeled preparation suitable to measure NMR spectrum. To overcome this obstacle, we tried to develop a over-production system of 15N-labeled P-glycoprotein using a methylotrophic yeast, Pichia pastoris. We obtained several ten milli-gram of purified preparation of 15N-labeled P-glycoprotein from a thermophilic eukaryote, Cyanidioschyzon merolae. We measured 1H-15N TROSY spectrum, suggesting normally folded protein behavior and the specimen is suitable for an analysis of dynamic structure.
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