title: Structure determination of a human virus by the combination of cryo-EM and X-ray crystallography.
abstract: 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.
authors.affiliation: Department of Biophysics, Health Science Centre, Peking University, Beijing, 100191 China.|Department of Biochemistry and Molecular Biophysics, Columbia University, New York, 10032 USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of BioSciences, Rice University, Houston, TX 77005 USA.
authors.forename: Tom S Y
authors.initials: TSY
authors.lastname: Guu
authors.affiliation: School of Life sciences, Sun Yat-sen University, Guangzhou, 510275 China.|State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, 510515 China.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Cao
authors.affiliation: School of Life sciences, Sun Yat-sen University, Guangzhou, 510275 China.
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Li
authors.affiliation: School of Life Science, Tsinghua University, Beijing, 100084 China.
authors.forename: Lingpeng
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Department of BioSciences, Rice University, Houston, TX 77005 USA.
authors.forename: Yizhi Jane
authors.initials: YJ
authors.lastname: Tao
authors.affiliation: School of Life sciences, Sun Yat-sen University, Guangzhou, 510275 China.
authors.forename: Jingqiang
authors.initials: J
authors.lastname: Zhang
keywords: Cryo-EM; Human virus; Structure determination; X-ray crystallography