Nanobiology Laboratories, Graduate School of Frontier Biosciences, Osaka University, Osaka 565-0871, Japan.
Proc Jpn Acad Ser B Phys Biol Sci. 2009;85(10):491-9. doi: 10.2183/pjab.85.491.
Until recently, actin was thought to act merely as a passive track for its motility partner, myosin, during actomyosin interactions. Yet a recent report having observed dynamical conformational changes in labeled skeletal muscle alpha-actin suggests that actin has a more active role. Because the labeling technique was still immature, however, conclusions regarding the significance of the different conformations are difficult to make. Here, we describe the preparation of fully active alpha-actin obtained from a baculovirus expression system. We developed alpha-actin recombinants, of which subdomains 1 and 2 have specific sites for fluorescent probes. This specific labeling technique offers to significantly expand the information acquired from actin studies.
直到最近，肌动蛋白被认为仅仅在肌球蛋白肌动蛋白相互作用过程中充当其运动伙伴的被动轨道。然而，最近的一份报告观察到标记的骨骼肌α-肌动蛋白的动态构象变化，表明肌动蛋白具有更积极的作用。然而，由于标记技术仍不成熟，因此很难得出关于不同构象意义的结论。在这里，我们描述了从杆状病毒表达系统获得的完全活性的α-肌动蛋白的制备方法。我们开发了α-肌动蛋白重组体，其中亚结构域 1 和 2 具有荧光探针的特定位点。这种特异性标记技术有望显著扩展从肌动蛋白研究中获得的信息。
