2009 Fiscal Year Final Research Report
Molecular Mechanism of energy transduction of dynein revealed by genetic engineering, structural and enzyme kinetics studies
| Project/Area Number |
17107003
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| Research Category |
Grant-in-Aid for Scientific Research (S)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biophysics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SUTOH Kazuo The University of Tokyo, 大学院・総合文化研究科, 教授 (20111453)
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| Co-Investigator(Renkei-kenkyūsha) |
KON Takahide 東京大学, 大学院・総合文化研究科, 助教 (30332620)
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| Project Period (FY) |
2005 – 2009
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| Keywords | 細胞質ダイニン / AAA+ファミリー / 微小管 / ATP加水分解酵素 / FRET / 電子顕微鏡 / パワーストローク |
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| Research Abstract |
Our major goal of this project is to elucidate the molecular mechanism of energy transduction of cytoplasmic dynein by genetic engineering, structural studies and ATPase kinetics analyses. Our major results are: (1) we have identified the minimal motor domain of cytoplasmic dynein and examined its structure by electron microscopy. (2) Based on ATPase kinetics, we have identified three ATPase sites and one ADP-binding site in the minimal motor domain (380kDa), among which one is the primary ATPase site and the others are regulatory sites. (3) We have identified intermediate steps of the ATPase cycle by means of stopped-flow kinetics. (4) We have discovered that the linker protruding from the core ring swings as a lever-arm at specific steps of the ATPase cycle of the primary ATPase site. (5) We have shown that structural changes of the microtubule-binding domain of dynein are induced at specific ATPase steps of the primary ATPase site. Based on these findings, we have proposed a molecular model of energy transduction of cytoplasmic dynein.
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[Journal Article] AAA+ Ring and Linker Swing Mechanism in the Dynein Motor2009
Author(s)
Anthony J. Roberts, Naoki Numata, Matt L. Walker, Yusuke S. Kato, Bara Malkova, Takahide Kon, Reiko Ohkura, Fumio Arisaka, Peter J. Knight, Kazuo Sutoh, Stan A. Burgess.
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Journal Title
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[Journal Article] Autonomous Loading/Unloading and Transport of Specified Cargoes Using DNA Hybridization and Biological Motor-Based Motility.2008
Author(s)
Hiyama, S., Inoue, T., Shima, T., Moritani, Y., Suda, T., Sutoh, K.
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Journal Title
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[Journal Article] A single-headed recombinant fragment of Dictyostelium cytoplasmic dynein can drive the robust sliding of microtubules.2004
Author(s)
Nishiura, M., Kon, T., Shiroguchi, K., Ohkura, R., Shima, T., Toyoshima, Y. Y., Sutoh, K.
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Journal Title
J.Biol.Chem. 279
Pages: 22799-22802
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