Hepatitis C Virus E2 Protein Ectodomain Is Essential for Assembly of Infectious Virions
Figure 3
Oxidation kinetics and glycosylation processing of E1E2 heterodimer. (a) S6.1 cells transfected with JFH1 or JFH1/Con1E2 RNA were pulse labeled with [35S] methionine and cysteine for 20 min and chased for different time periods, from 6 hrs to 48 hrs. PNSs were immunoprecipitated with a conformational monoclonal anti-E2 antibody (CBH2 or CBH5, resp., for genotype 2a and 1b) and analyzed on SDS-PAGE. Chase periods are reported above the lanes. Samples were analyzed on 10% SDS-PAGE under nonreducing and reducing conditions (DTT 200 mM). (b) Transfected cells were metabolically labeled for 6 hrs in presence of [35S] Met and Cys. E1 and E2 proteins were immunoprecipitated from the PNS with CBH-2 or CBH-5, respectively, for wt and chimeric species, treated () or not () with endo H and analyzed on 10% SDS-PAGE under nonreducing conditions. Note that Con1-derived proteins display an higher electrophoretic mobility than E1E2 proteins from JFH1 transfected cells. Symbol gly is for deglycosylated proteins.
