Department of Applied Physics, Graduate School of Engineering, Tohoku University, Sendai, Japan.
Department of Applied Physics, Graduate School of Engineering, Tohoku University, Sendai, Japan.
Biophys J. 2019 May 21;116(10):1952-1959. doi: 10.1016/j.bpj.2019.04.004. Epub 2019 Apr 11.
The bacterial flagellar motor is one of the most complex and sophisticated nanomachineries in nature. A duty ratio D is a fraction of time that the stator and the rotor interact and is a fundamental property to characterize the motor but remains to be determined. It is known that the stator units of the motor bind to and dissociate from the motor dynamically to control the motor torque depending on the load on the motor. At low load, at which the kinetics such as proton translocation speed limits the rotation rate, the dependency of the rotation rate on the number of stator units N implies D: the dependency becomes larger for smaller D. Contradicting observations supporting both the small and large D have been reported. A dilemma is that it is difficult to explore a broad range of N at low load because the stator units easily dissociate, and N is limited to one or two at vanishing load. Here, we develop an electrorotation method to dynamically control the load on the flagellar motor of Salmonella with a calibrated magnitude of the torque. By instantly reducing the load for keeping N high, we observed that the speed at low load depends on N, implying a small duty ratio. We recovered the torque-speed curves of individual motors and evaluated the duty ratio to be 0.14 ± 0.04 from the correlation between the torque at high load and the rotation rate at low load.
细菌鞭毛马达是自然界中最复杂和最精巧的纳米机械之一。占空比 D 是定子和转子相互作用的时间分数,是表征马达的基本特性,但仍有待确定。已知马达的定子单元动态地结合和离解马达,以根据马达上的负载来控制马达的扭矩。在低负载下,质子迁移速度等动力学限制了旋转速度,旋转速度对定子单元数量 N 的依赖性意味着 D:对于较小的 D,依赖性变得更大。已经报道了支持小 D 和大 D 的矛盾观察结果。一个难题是,由于定子单元容易离解,因此在低负载下很难探索广泛的 N,并且在负载消失时 N 限于一个或两个。在这里,我们开发了一种电动旋转方法,通过校准的扭矩幅度来动态控制沙门氏菌鞭毛马达的负载。通过立即降低负载以保持 N 高,我们观察到在低负载下速度取决于 N,这意味着占空比较小。我们恢复了单个马达的扭矩-速度曲线,并通过高负载下的扭矩与低负载下的旋转速度之间的相关性,评估占空比为 0.14±0.04。