Cryo-electron Microscopy of the Vacuolar ATPase Motor Reveals its Mechanical and Regulatory Complexity
Cryo-electron Microscopy of the Vacuolar ATPase Motor Reveals its Mechanical and Regulatory Complexity
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DOI：
10.1016/j.jmb.2009.01.014
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发表时间：
2009-03-06
影响因子：
5.6
通讯作者：
Harrison, Michael A.
中科院分区：
文献类型：
--
作者：
Muench, Stephen P.;Huss, Markus;Harrison, Michael A.
液泡H+-ATPase(V-ATPase)是一种由ATP驱动的旋转分子马达，是所有真核细胞中的跨膜质子泵。虽然它的活性是许多生理过程的基础，但我们对V-ATPase的结构和机制了解很少。利用烟草天牛(Manduca Sexta)酶的冷冻电子显微镜，我们计算了完整泵在其自然状态下的首次三维重建。16.5埃的分辨率明显高于已有的V-ATPase或相关的F1F0-ATPase的冷冻电子显微镜模型。连接V-1催化结构域和V-0膜结构域的四个茎结构的网络现在已经完全解析，表现出比F-ATPase中发现的更大的复杂性。三个外围的定子柄，将这些磁区连接到部分包围V-1和V-0之间区域的水平套圈。第四根柄是一个中心轴，它连接到V-0，但与V1的接触最少。几种亚单位晶体结构可以精确地用于重建。因此，该模型为领域之间的机械耦合和活动调节所涉及的关键组件的组织提供了新的见解。(C)2009爱思唯尔有限公司。保留所有权利。
The vacuolar H+-ATPase (V-ATPase) is an ATP-driven rotary molecular motor that is a transmembrane proton pump in all eukaryotic cells. Although its activity is fundamental to many physiological processes, our understanding of the structure and mechanism of the V-ATPase is poor. Using cryo-electron microscopy of the tobacco hornworm (Manduca sexta) enzyme, we have calculated the first 3D reconstruction of the intact pump in its native state. The resolution of 16.5 angstrom is significantly higher than that of previous cryo-electron microscopy models of either V-ATPase or the related F1F0-ATPase. A network of four stalk structures connecting the V-1 catalytic domain and the V-0 membrane domain is now fully resolved, demonstrating substantially greater complexity than that found in the F-ATPase. Three peripheral stator stalks, connect those domains to a horizontal collar that partly encircles the region between V-1 and V-0. The fourth stalk is a central axle that connects to V-0 but makes minimal contact with V1. Several subunit crystal structures can be fit accurately into the reconstruction. The model thus provides new insights into the organisation of key components involved in mechanical coupling between the domains and regulation of activity. (C) 2009 Elsevier Ltd. All rights reserved.