Expression, purification, and crystallization of HSV-1 glycoproteins for structure determination

Methods Mol Biol. 2014;1144:249-63. doi: 10.1007/978-1-4939-0428-0_17.

Abstract

HSV glycoproteins play important roles in the viral infectious cycle, particularly viral entry into the cell. Here we describe the protocol for expression, purification, and crystallization of viral glycoproteins based on those developed for the HSV-1 gB and HSV-2 gH/gL ectodomains. These protocols 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 other HSV glycoproteins for biochemical and structural studies.

MeSH terms

  • Cell Line
  • Crystallization
  • Gene Expression Regulation, Viral
  • Glycoproteins / chemistry*
  • Glycoproteins / genetics
  • Glycoproteins / isolation & purification
  • Herpesvirus 1, Human / chemistry
  • Herpesvirus 1, Human / genetics*
  • Herpesvirus 2, Human / chemistry*
  • Herpesvirus 2, Human / genetics
  • Humans
  • Molecular Biology / methods*
  • Protein Conformation

Substances

  • Glycoproteins