Molecular Structure and Function Program, The Hospital for Sick Children Research Institute, 686 Bay Street, Toronto, Ontario M5G 0A4, Canada.
Molecular Structure and Function Program, The Hospital for Sick Children Research Institute, 686 Bay Street, Toronto, Ontario M5G 0A4, Canada.; Department of Biochemistry, The University of Toronto, 1 King's College Circle, Toronto, Ontario M5S 1A8, Canada.; Department of Medical Biophysics, The University of Toronto, 101 College Street, Toronto, Ontario M5G 1L7, Canada.
Sci Adv. 2016 Jul 22;2(7):e1600725. doi: 10.1126/sciadv.1600725. eCollection 2016 Jul.
Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases.
电子冷冻显微镜（cryo-EM）极大地促进了我们对生物学中分子结构的理解。硬件和软件的最新创新使得 cryo-EM 成为一种可行的替代方法，适用于不适于 X 射线晶体学或核磁共振（NMR）光谱学的靶标。在某些情况下，cryo-EM 甚至成为首选方法，这些情况下 X 射线晶体学和 NMR 光谱学是可行的，但 cryo-EM 可以以更高的分辨率、更少的时间或更少的努力来确定结构。旋转腺苷三磷酸酶（ATPases）对于维持细胞内稳态至关重要。这些酶分别将腺苷三磷酸的合成或水解与跨膜电化学离子梯度的利用或产生偶联。然而，完整旋转 ATPase 的膜嵌入性质和构象异质性阻止了它们迄今为止的高分辨率结构分析。最近 cryo-EM 方法在不同类型的旋转 ATPase 中的应用导致了对这些酶的结构和功能的理解的突然进展，揭示了显著的构象异质性和特征性的跨膜α螺旋，这些螺旋相对于膜高度倾斜。在这篇综述中，我们将讨论最近关于旋转 ATPase 结构和功能的研究进展，特别关注液泡型 ATPase。
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