Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination.
Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination.
复制标题
DOI：
10.1007/978-1-4939-9814-2_23
复制
发表时间：
2019-10
影响因子：
--
通讯作者：
E. White;S. Stampfer;E. Heldwein
中科院分区：
文献类型：
--
作者：
E. White;S. Stampfer;E. Heldwein
单纯疱疹病毒利用病毒包膜上显示的糖蛋白在病毒感染周期中执行各种功能。对这些病毒糖蛋白的结构和功能的研究可以从对纯化蛋白的生化、生物物理和结构分析中受益。在此，我们描述了一种从昆虫细胞中表达和纯化病毒糖蛋白的通用方案，该方案基于针对HSV-1 gB和HSV-2 Gh/gl胞外区开发的糖蛋白，以及这些糖蛋白的结晶方案。该方法可用于产生毫克量的野生型(WT)或突变型gB和Gh/gl胞外区，或可用于从HSV或其他疱疹病毒产生纯化的糖蛋白胞外区，用于生化和结构研究。
Herpes simplex viruses utilize glycoproteins displayed on the viral envelope to perform a variety of functions in the viral infectious cycle. Structural and functional studies of these viral glycoproteins can benefit from biochemical, biophysical, and structural analysis of purified proteins. Here, we describe a general protocol for expression and purification of viral glycoproteins from insect cells based on those developed for the HSV-1 gB and HSV-2 gH/gL ectodomains as well as the protocol for crystallization of these glycoproteins. This protocol can be used for generating milligram amounts of wild-type (WT) or mutant gB and gH/gL ectodomains or can be adapted to produce purified ectodomains of glycoproteins from HSV or other herpesviruses for biochemical and structural studies.