Measuring Hepatitis C Virus Envelopment by Using a Proteinase K Protection Assay
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.
Key wordsHepatitis C virus Envelopment Proteinase K Immunoblot Electroporation Protection assay
Research on hepatitis C virus in the Horner laboratory is supported by funds from the National Institutes of Health (NIH) (R01AI125416 and R21AI124100), a Duke School of Medicine Whitehead Scholarship, and the Burroughs Wellcome Fund. AER has received support from NIH 1F31AI131477-01A1 and T32CA009111.