| Project/Area Number |
18570153
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | Osaka University |
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Principal Investigator |
IWANE Atsuko Osaka University, Graduate School of Frontier Biosciences, Associate Professor (30252638)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Single molecule techniques / Molecular machine / Molecular motor / Protein / Biotechnology |
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| Research Abstract |
It is likely to be able to explain as a movement mechanism of Myosin by "biased brownian movement". We showed the hypothesis to which we say that "hot spot" was raised on the actin filament during acto-myosin interaction in our paper in 2002, but I'd like to take out more evidence which can be the conclusive evidence which moves in one direction.
First of all, as this result of two years 1) We obtained a preliminary similar result, the confirmation of the hot spots on the actin filament during actomyosin interaction with a Cryo-electron microscope under a more physiological condition. And then, 2) We succeeded in observation of a single molecule movement of the myosin using the myosin Va by which one after Quantum Dot addition maintains a way sufficiently as well as GFP fused myosin Va, VI and moved prossecive steps which are 75 nm averagely on actin filament. I succeeded to catch the scene directly where I get one-directional movement when I added Cy3-ATP to this system, to do simultaneous measurement with ATP hydrolysis and a step. In addition, 3) we clarified the evidence an actin filament rotate at the twice ATP dependent 90 degrees per one walking step of myosin Va. I'm thinking rotative direction revolve using an Brownian motion at randam motion. 4) We confirmed an intramolecular conformational change in actin using a single molecule FRET to visualize the activation state of it. We succeeded in detection of an intramolecular conformational fluctuation state who suggest the activation state of the actin. Establishment of recombinant actin is the important research achievement from which I'll get much information from now on. Finally, 5) I obtained results about the role of the neck domain of myosin and the common characteristic of the acto-myosin interaction mechanism on the actin filament.
The research achievements (1,2) are getting ready as some papers at present. (3-5) had been reported as many oral presentations at many meeting and theses of art and science.
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