2013 Fiscal Year Final Research Report
Affinity regulation mechanism of microtubule-binding domain of cytoplasmic dynein
| Project/Area Number |
24770090
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
SHIMADA Ichio 東京大学, 大学院薬学系研究科 (70196476)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Keywords | 核磁気共鳴法 / クライオ電顕 / モーター蛋白質 |
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| Research Abstract |
Cytoplasmic dynein is a motor protein that walks on microtubules (MTs). The microtubule-binding domain (MTBD) is located at the tip of the anti-parallel coiled-coil, which is protruded from the ATPase domain of dynein heavy chain. It has been considered that the change in the association mode of the coiled-coil alters the MT affinity of the MTBD. However, the structural mechanism underlying the affinity regulation of the MTBD remains elusive.
Here, we designed two MTBD constructs, termed MTBD-High and MTBD-Low, which are stabilized either in the high or low affinity state for MTs, respectively, by introducing a disulfide bond between the coiled-coil helix. We performed NMR analyses of MTBD-High and MTBD-Low, in order to elucidate differences in the structure and the MT-binding mode in the high and low affinity states. Based on NMR data in combination with the cryoEM analysis of the MTBD-High/MTs complex, we modeled the complex structure of the MTBD and MTs in the strong binding state.
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