Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Capsid / ultrastructure*
  • Capsid Proteins / chemistry
  • Capsid Proteins / ultrastructure*
  • Cryoelectron Microscopy
  • DNA Packaging / genetics
  • Herpesvirus 1, Human / chemistry
  • Herpesvirus 1, Human / genetics*
  • Humans
  • Image Processing, Computer-Assisted
  • Molecular Biology / methods*

Substances

  • Capsid Proteins