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  • The hepatitis C virus E1 glycoprotein undergoes productive folding but accelerated degradation when expressed as an individual subunit in CHO cells.

The hepatitis C virus E1 glycoprotein undergoes productive folding but accelerated degradation when expressed as an individual subunit in CHO cells.

PloS one (2011-08-23)
Valentina Botti, Alessia Bianchi, Steven K H Foung, Marcello Merola
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Endoglycosidase F3 from Elizabethkingia miricola, recombinant, expressed in E. coli, 30 U/mg
Sigma-Aldrich
Endoglycosidase H from Streptomyces plicatus, recombinant, expressed in E. coli, buffered aqueous solution
Sigma-Aldrich
Endoglycosidase F1 from Elizabethkingia miricola, recombinant, expressed in E. coli, ≥16 U/mg, buffered aqueous solution
Sigma-Aldrich
Endoglycosidase F2 from Elizabethkingia miricola, recombinant, expressed in E. coli, 20 U/mg