Expression, Purification, and Crystallization of Full-Length HSV-1 gB for Structure Determination

Methods Mol Biol. 2020:2060:395-407. doi: 10.1007/978-1-4939-9814-2_24.

Abstract

HSV glycoproteins play important roles in the viral life cycle, particularly viral cell entry. Here we describe the protocol for expression, purification, and crystallization of full-length HSV-1 glycoprotein B. The protocol provides a framework for incorporating transmembrane domain-stabilizing amphipols into the crystallization setup and can be adapted to isolate other complete HSV glycoproteins.

Keywords: Crystallography; Ectodomain; Glycoproteins; Herpes simplex viruses; Membrane protein; Protein purification; Viral entry.

MeSH terms

  • Animals
  • Crystallography, X-Ray
  • Gene Expression*
  • Herpesvirus 1, Human* / chemistry
  • Herpesvirus 1, Human* / genetics
  • Herpesvirus 1, Human* / metabolism
  • Protein Domains
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Sf9 Cells
  • Spodoptera
  • Viral Envelope Proteins* / biosynthesis
  • Viral Envelope Proteins* / chemistry
  • Viral Envelope Proteins* / genetics
  • Viral Envelope Proteins* / isolation & purification

Substances

  • Recombinant Proteins
  • Viral Envelope Proteins
  • glycoprotein B, Simplexvirus