Role of actin potential gradient as to the acto-myosin motility mechanism

这可能可以解释为肌球蛋白的一种“偏布朗运动”的运动机制。我们在2002年的论文中提出了在肌动蛋白-肌球蛋白相互作用过程中肌动蛋白丝上产生“热点”的假设，但我想拿出更多的证据，这是一个方向上的结论性证据。首先，作为这两年的结果1)我们获得了一个初步的类似结果，在更生理的条件下，用冷冻电镜确认了肌动球蛋白相互作用过程中肌动蛋白丝上的热点。然后，2)利用肌凝蛋白Va成功观察到肌凝蛋白的单分子运动，其中量子点加入后的肌凝蛋白Va、VI充分保持了一种方式，GFP融合肌凝蛋白Va、VI并在肌动蛋白丝上移动了平均75 nm的前进步。当我将Cy3-ATP加入到这个体系中，我成功地直接捕捉到了我得到单向运动的场景，用ATP水解和一步同时测量。此外，3)我们澄清了肌动蛋白丝以两次ATP依赖的90度旋转的证据，每走一步肌凝蛋白Va。我认为旋转方向以随机运动的布朗运动旋转。4)我们使用单分子FRET证实了肌动蛋白的分子内构象变化，以可视化其激活状态。我们成功地检测到分子内构象波动状态，提示肌动蛋白的激活状态。重组肌动蛋白的建立是一项重要的研究成果，今后我将从中获得很多信息。最后，5)获得了肌动蛋白颈域的作用以及肌动蛋白-肌动蛋白相互作用机制的共同特点。研究成果（1、2）目前正在准备发表论文。（3-5）在许多会议上作了口头报告，并在艺术和科学领域发表了论文。

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.