Department of Cell Biology, Graduate School of Medicine, the University of Tokyo, Tokyo, Japan.
Department of Cell Biology, Graduate School of Medicine, the University of Tokyo, Tokyo, Japan; Department of Physics, Graduate School of Science, the University of Tokyo, Tokyo, Japan; Universal Biology Institute and International Research Center for Neurointelligence, the University of Tokyo, Tokyo, Japan; Laboratory for Cell Polarity Regulation, Center for Biosystems Dynamics Research (BDR), RIKEN, Osaka, Japan.
Biophys J. 2024 Oct 15;123(20):3587-3599. doi: 10.1016/j.bpj.2024.09.010. Epub 2024 Sep 11.
Many bacteria enable locomotion by rotating their flagellum. It has been suggested that this rotation is realized by the rotary motion of the stator unit, MotAB, which is driven by proton transfer across the membrane. Recent cryo-electron microscopy studies have revealed a 5:2 MotAB configuration, in which a MotB dimer is encircled by a ring-shaped MotA pentamer. Although the structure implicates the rotary motion of the MotA wheel around the MotB axle, the molecular mechanisms of rotary motion and how they are coupled with proton transfer across the membrane remain elusive. In this study, we built a structure-based computational model for Campylobacter jejuni MotAB, conducted comprehensive protonation-state-dependent molecular dynamics simulations, and revealed a plausible proton-transfer-coupled rotation pathway. The model assumes rotation-dependent proton transfer, in which proton uptake from the periplasmic side to the conserved aspartic acid in MotB is followed by proton hopping to the MotA proton-carrying site, followed by proton export to the CP. We suggest that, by maintaining two of the proton-carrying sites of MotA in the deprotonated state, the MotA pentamer robustly rotates by ∼36° per proton transfer across the membrane. Our results provide a structure-based mechanistic model of the rotary motion of MotAB in bacterial flagellar motors and provide insights into various ion-driven rotary molecular motors.
许多细菌通过旋转鞭毛来实现运动。有人提出,这种旋转是通过定子单元 MotAB 的旋转运动来实现的,MotAB 由质子穿过膜的转移驱动。最近的低温电子显微镜研究揭示了一种 5:2 的 MotAB 结构,其中一个 MotB 二聚体被一个环形的 MotA 五聚体包围。虽然该结构暗示了 MotA 轮围绕 MotB 轴的旋转运动,但旋转运动的分子机制以及它们与质子穿过膜的转移如何偶联仍然难以捉摸。在这项研究中,我们为弯曲杆菌 MotAB 构建了一个基于结构的计算模型,进行了全面的质子化状态依赖的分子动力学模拟,并揭示了一种合理的质子转移偶联旋转途径。该模型假设旋转依赖的质子转移,其中从周质侧到 MotB 中的保守天冬氨酸摄取质子,然后质子跳跃到 MotA 的质子携带位点,然后质子被运送到 CP。我们认为,通过保持 MotA 中两个质子携带位点处于去质子化状态,MotA 五聚体可以通过每轮质子穿过膜的转移稳定地旋转约 36°。我们的结果为细菌鞭毛马达中 MotAB 的旋转运动提供了一个基于结构的机制模型,并为各种离子驱动的旋转分子马达提供了深入的了解。