Large-Scale Expression and Purification of Mumps Virus Hemagglutinin-Neuraminidase for Structural Analyses and Glycan-Binding Assays

Methods Mol Biol. 2020:2132:641-652. doi: 10.1007/978-1-0716-0430-4_55.

Abstract

Many viruses utilize cell-surface glycans as receptors for host cell entry. Viral surface glycoproteins specifically interact with glycan motifs, which strongly contributes to viral tropism. Recently, the interactions between host cell glycan receptors and the mumps virus (MuV) hemagglutinin-neuraminidase (MuV-HN) protein were characterized by determining the co-crystal structure of MuV-HN in complex with glycan receptors. Here, we describe protocols for large-scale expression, purification and crystallization of MuV-HN proteins for structural analyses and glycan-binding assays with the overarching goal of investigating glycan-protein interactions.

Keywords: Glycan receptors; Glycoprotein; HEK293S GnTI(−) cells; Hemagglutinin-neuraminidase; Large-scale protein expression; Mumps virus; Purification; Structure; X-ray crystallography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Affinity
  • Chromatography, Gel
  • Crystallography, X-Ray
  • HEK293 Cells
  • HN Protein / chemistry*
  • HN Protein / isolation & purification
  • HN Protein / metabolism*
  • Humans
  • Mumps / virology*
  • Mumps virus / physiology*
  • N-Acetylglucosaminyltransferases / deficiency
  • Polysaccharides / metabolism*
  • Protein Binding
  • Protein Domains
  • Protein Engineering
  • Viral Tropism
  • Virus Internalization

Substances

  • HN Protein
  • Polysaccharides
  • N-Acetylglucosaminyltransferases
  • alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I