title: Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components.
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.
authors.affiliation: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Ryan H
authors.initials: RH
authors.lastname: Rochat
authors.forename: Corey W
authors.initials: CW
authors.lastname: Hecksel
authors.forename: Wah
authors.initials: W
authors.lastname: Chiu
mesh_terms: D002213:Capsid / Q000648:ultrastructure*; D036022:Capsid Proteins / Q000737:chemistry / Q000648:ultrastructure*; D020285:Cryoelectron Microscopy; D042003:DNA Packaging / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008967:Molecular Biology / Q000379:methods*
chemical_list: D036022:Capsid Proteins