Structure determination of a human virus by the combination of cryo-EM and X-ray crystallography.
Structure determination of a human virus by the combination of cryo-EM and X-ray crystallography.
复制标题
DOI：
10.1007/s41048-016-0027-2
复制
发表时间：
2016
影响因子：
--
通讯作者：
Zhang, Jingqiang
中科院分区：
文献类型：
--
作者：
Liu, Zheng;Guu, Tom S Y;Cao, Jianhao;Li, Yinyin;Cheng, Lingpeng;Tao, Yizhi Jane;Zhang, Jingqiang
病毒的3D原子结构为我们了解病毒的生命周期和抗病毒药物的开发提供了见解。X射线晶体学和冷冻电镜已被用于确定病毒的原子结构。然而，生物样品的有限可用性、由于病毒感染引起的生物安全问题以及有时病毒的固有特性，在确定病毒结构时结合两种方法造成困难。这些使得解决一些医学上重要的病毒的高分辨率结构非常具有挑战性。在这里，我们描述了我们最近采用的协议，用于确定高分辨率的病毒样颗粒戊型肝炎病毒（HEV），病毒性肝炎的病原体在人类的结构。这些方案包括利用重组杆状病毒系统产生足够量的病毒颗粒，单颗粒冷冻-EM以获得中间分辨率结构作为定相模型，以及X射线晶体学用于最终的原子结构测定。我们的方案已经解决了戊型肝炎病毒的结构分辨率为3.5 μ m。该组合方法通常适用于其他人类感染性病毒。
Virus 3D atomic structures provide insight into our understanding of viral life cycles and the development of antiviral drugs. X-ray crystallography and cryo-EM have been used to determine the atomic structure of viruses. However, limited availability of biological samples, biosafety issues due to virus infection, and sometimes inherent characteristics of viruses, pose difficulties on combining both methods in determining viral structures. These have made solving the high resolution structure of some medically important viruses very challenging. Here, we describe our recently employed protocols for determining the high-resolution structure of the virus-like particle of hepatitis E virus (HEV), a pathogen of viral hepatitis in human. These protocols include utilizing recombinant baculovirus system to generate sufficient amount of virus particles, single-particle cryo-EM to get an intermediate resolution structure as a phasing model, and X-ray crystallography for final atomic structure determination. Our protocols have solved the hepatitis E virus structure to the resolution of 3.5 Å. The combined methodology is generally applicable to other human infectious viruses.