| Project/Area Number |
15K14463
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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 |
Structural biochemistry
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| Research Institution | Tokyo Metropolitan University |
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Principal Investigator |
Mishima Masaki 首都大学東京, 理工学研究科, 准教授 (70346310)
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| Co-Investigator(Kenkyū-buntansha) |
白井 康仁 神戸大学, 農学研究科, 教授 (60263399)
伊藤 隆 首都大学東京, 理工学研究科, 教授 (80261147)
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| Project Period (FY) |
2015-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | プロテインライゲーション / NMR / マルチドメインタンパク質 / キナーゼ / sortase |
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| Outline of Final Research Achievements |
It is generally recognized that kinases consist of multiple domain (mutli-domain proteins), which have flexible linker(s). This flexibility may hamper crystallization. In this study, our purpose is visualization of structural rearrangements of multi-domain kinases (transition from inhibited inactive states to activated states) using solution techniques at molecular level to understand the regulation mechanism. We have succeeded in domain ligation of C1B domain with kinase domain of protein kinase C using sortase. Further, we synthesized DOTA-M8, a rigid chelator for lanthanide ions, to measure PCS to obtain long-range distance information. It contains maleimide group which is useful to attach to proteins. Using this chelator, we actually measured PCS.
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