用于解析单纯疱疹病毒1型衣壳相关成分结构的冷冻电镜技术。
Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components.
作者信息
Rochat Ryan H, Hecksel Corey W, Chiu Wah
机构信息
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
出版信息
Methods Mol Biol. 2014;1144:265-81. doi: 10.1007/978-1-4939-0428-0_18.
Abstract
Electron cryo-microscopy has become a routine technique to determine the 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 α-helices and β-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.
摘要
电子冷冻显微镜已成为确定生物化学纯化的单纯疱疹病毒衣壳颗粒结构的常规技术。本章描述了通过冷冻保存制备标本的程序；在电子冷冻显微镜中进行低剂量和低温成像；以及用于重建的数据处理。这种方法已经获得了二十面体衣壳壳的亚纳米分辨率结构，其中可以识别单个亚基的α螺旋和β折叠。在重建步骤中对称性的放宽使我们能够解析位于衣壳12个顶点之一的DNA包装蛋白。
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