Boschert Ryan, Adler Frederick R, Blair David F
Department of Biology, University of Utah, Salt Lake City, UT 84112.
Department of Biology, University of Utah, Salt Lake City, UT 84112
Proc Natl Acad Sci U S A. 2015 Apr 14;112(15):4755-60. doi: 10.1073/pnas.1419955112. Epub 2015 Mar 30.
Physiological properties of the flagellar rotary motor have been taken to indicate a tightly coupled mechanism in which each revolution is driven by a fixed number of energizing ions. Measurements that would directly test the tight-coupling hypothesis have not been made. Energizing ions flow through membrane-bound complexes formed from the proteins MotA and MotB, which are anchored to the cell wall and constitute the stator. Genetic and biochemical evidence points to a "power stroke" mechanism in which the ions interact with an aspartate residue of MotB to drive conformational changes in MotA that are transmitted to the rotor protein FliG. Each stator complex contains two separate ion-binding sites, raising the question of whether the power stroke is driven by one, two, or either number of ions. Here, we describe simulations of a model in which the conformational change can be driven by either one or two ions. This loosely coupled model can account for the observed physiological properties of the motor, including those that have been taken to indicate tight coupling; it also accords with recent measurements of motor torque at high load that are harder to explain in tight-coupling models. Under loads relevant to a swimming cell, the loosely coupled motor would perform about as well as a two-proton motor and significantly better than a one-proton motor. The loosely coupled motor is predicted to be especially advantageous under conditions of diminished energy supply, or of reduced temperature, turning faster than an obligatorily two-proton motor while using fewer ions.
鞭毛旋转马达的生理特性表明存在一种紧密偶联机制,即每一次旋转由固定数量的供能离子驱动。尚未进行能直接验证紧密偶联假说的测量。供能离子流经由MotA和MotB蛋白形成的膜结合复合物,MotA和MotB锚定在细胞壁上,构成定子。遗传学和生物化学证据指向一种“动力冲程”机制,即离子与MotB的一个天冬氨酸残基相互作用,驱动MotA的构象变化,并传递给转子蛋白FliG。每个定子复合物包含两个独立的离子结合位点,这就引发了一个问题:动力冲程是由一个、两个还是任意数量的离子驱动的。在这里,我们描述了一个模型的模拟,其中构象变化可以由一个或两个离子驱动。这个松散偶联模型可以解释观察到的马达生理特性,包括那些被认为表明紧密偶联的特性;它也与最近在高负载下对马达扭矩的测量结果相符,而在紧密偶联模型中这些结果更难解释。在与游动细胞相关的负载条件下,松散偶联马达的表现与双质子马达相当,且明显优于单质子马达。预计在能量供应减少或温度降低的条件下,松散偶联马达将特别有利,它比强制双质子马达转得更快,同时消耗的离子更少。