Laboratory of Molecular Physiology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA.
Biophys J. 2009 Dec 16;97(12):3123-31. doi: 10.1016/j.bpj.2009.09.033.
Myosin V is a single-molecule motor that moves organelles along actin. When myosin V pulls loads inside the cell in a highly viscous environment, the force on the motor is unlikely to be constant. We propose that the tether between the single-molecule motor and the cargo (i.e., the extended tail domain of the molecule) must be able to absorb the sudden mechanical motions of the motor and allow smooth relaxation of the motion of the cargo to a new position. To test this hypothesis, we compared the elastic properties of the extended tail domains of processive (mouse myosin Va) and nonprocessive (Drosophila myosin V) molecular motors. The extended tail domain of these myosins consists of mechanically strong coiled-coil regions interspersed with flexible loops. In this work we explored the mechanical properties of coiled-coil regions using atomic force microscopy. We found that the processive and nonprocessive coiled-coil fragments display different unfolding patterns. The unfolding of coiled-coil structures occurs much later during the atomic force microscopy stretch cycle for processive myosin Va than for nonprocessive Drosophila myosin V, suggesting that this elastic tether between the cargo and motor may play an important role in sustaining the processive motions of this single-molecule motor.
肌球蛋白 V 是一种沿肌动蛋白移动细胞器的单分子马达。当肌球蛋白 V 在高粘性环境中拉动细胞内的负载时,作用在马达上的力不太可能是恒定的。我们提出,单分子马达和货物(即分子的延伸尾部域)之间的系绳必须能够吸收马达的突然机械运动,并允许货物的运动平稳地松弛到新的位置。为了验证这一假设,我们比较了有向(小鼠肌球蛋白 Va)和无向(果蝇肌球蛋白 V)分子马达的延伸尾部域的弹性特性。这些肌球蛋白的延伸尾部域由机械强度高的卷曲螺旋区和柔性环组成。在这项工作中,我们使用原子力显微镜探索了卷曲螺旋区的力学性质。我们发现,有向和无向卷曲螺旋片段显示出不同的展开模式。在原子力显微镜拉伸循环中,有向肌球蛋白 Va 的卷曲螺旋结构的展开发生得比无向果蝇肌球蛋白 V 晚得多,这表明货物和马达之间的这种弹性系绳可能在维持这种单分子马达的有向运动中发挥重要作用。