Analysis of a ADP-bound non-muscle myosin molecule

有报道称，在肌动球蛋白复合体中加入ADP后，平滑肌肌凝蛋白和刷状边界肌凝蛋白的轻链部分发生构象变化。光镊实验表明，力的产生分两步进行，从而提出了第二步力是由ADP解离产生的假设。然而，未观察到添加ADP对平滑肌力的降低。因此，ADP在战斗链中看到的构象变化被认为是菌株依赖的ADP解离，而不是力量冲程的一部分。本研究的目的是阐明菌株依赖的ADP解离。遗憾的是，由于实验装置和方法的研制花费了大量的时间，因此没有得到菌株依赖性ADP解离的结论。然而，我们已经制作了一个精细的仪器来测量从刷状边界产生的单个肌球蛋白分子的力。我们描述了装置的研制过程、刷边肌球蛋白的制备过程、微球样品的制备过程，并展示了单个肌球蛋白分子的产力结果。

It has been reported that conformational changes occurred at the light chain portion of smooth muscle myosin and brush border myosin when ADP was added to the actomyosin complex. And the optical tweezer experiment has revealed that the foece generation occurred by two steps, leading the hypothesis that the second step of force is generated by the ADP dissociation. However, the decrease in the force of smooth muscle by adding ADP was not observed. So, the conformational changes seen in the fight chain by ADP is thought to be the strain-dependent ADP dissociation, not a portion of a power stroke. The purpose of this study was to elucidate the strain-dependent ADP dissocation. Unfortunately, a lot of time was spent to develope the experimental apparatus and the method, so the conclusion on the strain-dependent ADP dissociation was not obtained. However, we have made a fine apparatus that measure the force generation of a single myosin molecule from brush border.We describe the process of developing the apparatus, the preparation of the brush border myosin, and the preparation of the beads sample, and show the result of the force generation of single myosin molecule.