Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components.
Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components.
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DOI：
10.1007/978-1-4939-0428-0_18
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Chiu, Wah
中科院分区：
文献类型：
--
作者：
Rochat, Ryan H;Hecksel, Corey W;Chiu, Wah
电子冷冻显微镜已成为测定生化纯化的单纯疱疹病毒衣壳颗粒结构的常规技术。本章描述了冷冻保存标本的制备过程；低温电镜低剂量成像以及用于重建的数据处理。这种方法已经产生了亚纳米分辨率的二十面体衣壳结构，其中可以识别单个亚基的α螺旋和β片。重构步骤中对称性的松弛使我们能够解析位于衣壳12个顶点之一的DNA包装蛋白。
Electron cryo-microscopy has become a routine technique to determine structure of biochemically purified herpes simplex virus capsid particles. This chapter describes the procedures of specimen preparation by cryopreservation; low dose and low temperature imaging in an electron cryo-microscope; and data processing for reconstruction. This methodology has yielded subnanometer resolution structures of the icosahedral capsid shell where alpha helices and beta sheets of individual subunits can be recognized. A relaxation of the symmetry in the reconstruction steps allows us to resolve the DNA packaging protein located at one of the 12 vertices in the capsid.