Single-particle Cryo-EM of calcium release channels: structural validation

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• 作者：Steven J Ludtke ; Irina I Serysheva
• 刊名：Current Opinion in Structural Biology
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：23
• 期：5
• 页码：755-762
• 全文大小：1698 K

文摘

Few tools are available to determine the structure of large integral membrane proteins such as intracellular Ca²⁺ release channels, RyRs and IP₃Rs. Single particle cryo-EM can readily determine the structure of such channels to intermediate resolution, and can be used to quantitatively assess conformational variability. However, due to the, often low, image contrast of these cryospecimens, methods for validation are critical to insure the accuracy of such structures, and to put limits on their interpretability. The low-resolution structure of RyR has been well established for some time, but high-resolution has been slow to emerge. The structure of IP₃R channel by cryo-EM had a number of false-starts, but improved validation methods have recently lead to a demonstrably accurate reconstruction.