Measuring Hepatitis C Virus Envelopment by Using a Proteinase K Protection Assay

Methods Mol Biol. 2019:1911:209-217. doi: 10.1007/978-1-4939-8976-8_14.

Abstract

The infectious virion of hepatitis C virus (HCV) is made up of the viral nucleocapsid surrounded by an envelope that contains an ER-derived membrane bilayer, cellular lipids, and the viral E1 and E2 glycoproteins. Because the infectious HCV particle contains both protein and lipid layers, selective disruption of these layers and analysis for the presence or absence of resulting virion components can be used to study the virion assembly process. This chapter describes an experimental method to measure HCV virion envelopment, which can reveal the mechanisms of how specific viral protein-protein interactions and host factors contribute to the process of HCV envelopment.

Keywords: Electroporation; Envelopment; Hepatitis C virus; Immunoblot; Protection assay; Proteinase K.

Publication types

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

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Line
  • Electroporation / methods
  • Endopeptidase K / metabolism*
  • Hepacivirus / genetics
  • Hepacivirus / physiology*
  • Hepatitis C / virology*
  • Humans
  • Immunoblotting / methods
  • RNA, Viral / genetics
  • Viral Core Proteins / genetics
  • Viral Core Proteins / metabolism*
  • Virion / genetics
  • Virion / physiology*
  • Virus Assembly

Substances

  • RNA, Viral
  • Viral Core Proteins
  • Endopeptidase K