title: The hepatitis C virus E1 glycoprotein undergoes productive folding but accelerated degradation when expressed as an individual subunit in CHO cells.
abstract: Hepatitis C Virus E1E2 heterodimers are components of the viral spike. Although there is a general agreement on the necessity of the co-expression of both E1 and E2 on a single coding unit for their productive folding and assembly, in a previous study using an in vitro system we obtained strong indications that E1 can achieve folding in absence of E2. Here, we have studied the folding pathway of unescorted E1 from stably expressing CHO cells, compared to the folding observed in presence of the E2 protein. A DTT-resistant conformation is achieved by E1 in both situations, consistent with the presence of an E2-independent oxidative pathway. However, while the E1E2 heterodimer is stable inside cells, E1 expressed alone is degraded within a few hours. On the other hand, the oxidation and stability of individually expressed E2 subunits is dependent on E1 co-expression. These data are consistent with E1 and E2 assisting each other for correct folding via different mechanisms: E2 assists E1 by stabilizing a semi-native conformation meanwhile E1 drives E2 towards a productive folding pathway.
authors.affiliation: Novartis Vaccines and Diagnostics, Siena, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Botti
authors.forename: Alessia
authors.initials: A
authors.lastname: Bianchi
authors.forename: Steven K H
authors.initials: SK
authors.lastname: Foung
authors.forename: Marcello
authors.initials: M
authors.lastname: Merola
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004229:Dithiothreitol / Q000737:chemistry / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D047468:Immunoprecipitation; D007700:Kinetics; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D010084:Oxidation-Reduction; D011487:Protein Conformation / Q000187:drug effects; D017510:Protein Folding; D055503:Protein Multimerization; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: C087035:E1 protein, Hepatitis C virus; D021122:Protein Subunits; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D004229:Dithiothreitol