2013 Fiscal Year Final Research Report
Does dynein generate force by the linker swing?
| Project/Area Number |
23370075
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Waseda University |
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Principal Investigator |
SUTOH Kazuo 早稲田大学, 理工学術院, 教授 (20111453)
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| Co-Investigator(Renkei-kenkyūsha) |
KINOSITA Kazuhiko 早稲田大学, 理工学術院, 教授 (30124366)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Keywords | 生物物理 / 運動・輸送 / 1分子計測 / 構造解析 / ダイニン |
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| Research Abstract |
The question addressed here is whether dynein generates force by ATPase-dependent linker swing. The following system has been set up for observing the linker swing at a single molecule level. The motor domain was attached on glass surface through interactions between His-tags and anti-His-tag antibodies. A small bead was attached at the end of the linker through the avidin-biotin interaction. Beads motions were observed in the presence of ATP under a microscope. Some beads (~0.1%) showed ~90 degree rotation as expected for the ATPase-dependent linker swing. To check if these rotations correspond to linker swings, ATP was quickly exchanged to AMPPNP,which should stop the linker swing. Each rotating beads were checked immediately after the solution exchange. Rotation of each bead examined so far did not stop. Further data collection is required to pick up beads motions reflecting the linker swing.
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