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

用“偏布朗运动”来解释肌球蛋白的运动机制是可能的。我们在2002年的论文中提出了我们所说的在肌动蛋白-肌球蛋白相互作用期间在肌动蛋白丝上产生“热点”的假设，但我想拿出更多的证据，这些证据可以成为朝着一个方向移动的决定性证据。首先，作为两年的结果1）我们获得了初步的类似结果，在更生理条件下，用冷冻电子显微镜证实肌动球蛋白相互作用期间肌动蛋白丝上的热点。然后，2）我们成功地观察到了肌球蛋白Va的单分子运动，通过该运动，加入量子点后的一个肌球蛋白Va与GFP融合的肌球蛋白Va、VI一样充分地保持了一种方式，并在肌动蛋白丝上移动了平均75 nm的连续步骤。当我将Cy 3-ATP加入到这个系统中时，我成功地直接捕捉到了我得到单向运动的场景，以进行ATP水解和一步的同时测量。此外，我们还阐明了肌球蛋白Va每走一步，肌动蛋白丝以ATP依赖的90度旋转两次的证据。我在考虑旋转方向的旋转，用布朗运动和兰登运动。4)我们用单分子FRET技术观察了肌动蛋白的活化状态，证实了肌动蛋白分子内的构象变化，并成功地检测到了肌动蛋白分子内的构象波动状态，表明了肌动蛋白的活化状态。重组肌动蛋白的建立是重要的研究成果，我将从中获得更多信息。最后，5）我获得了有关肌球蛋白颈区的作用以及肌动蛋白丝上肌动蛋白-肌球蛋白相互作用机制的共同特征的结果。研究成果（1，2）目前正在准备一些论文。(3-5)在许多会议上作口头报告，并发表艺术和科学论文。

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.