2016 Fiscal Year Final Research Report
Dynamical study of multi-domain kinases reconstructed by domain ligation
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 |
Research Field |
Structural biochemistry
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Research Institution | Tokyo Metropolitan University |
Principal Investigator |
Mishima Masaki 首都大学東京, 理工学研究科, 准教授 (70346310)
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Project Period (FY) |
2015-04-01 – 2017-03-31
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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 |
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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Free Research Field |
構造生物化学
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