Hepatitis C virus glycoprotein folding: disulfide bond formation and association with calnexin.
The hepatitis C virus (HCV) glycoproteins (E1 and E2) are released from the polyprotein by signal peptidase-mediated cleavage and interact to form a heterodimer. Since properly folded subunits are usually required for specific recognition and stable oligomer formation, the rate of stable E1E2 complex formation, which is low, may be limited by the rate of HCV E1 and/or E2 folding. In this study, the folding of the HCV E1 and E2 glycoproteins was monitored by observing the kinetics of intramolecular disulfide bond formation. The association/dissociation of E1 and E2 with calnexin was also examined, since this molecular chaperone appears to play a major role in quality control via retention of incompletely folded or misfolded proteins in the endoplasmic reticulum. Our results indicate that the disulfide-dependent folding of E2 occurs rapidly and appears to be complete upon cleavage of the precursor E2-NS2. In contrast, folding of E1 is slow (> 1 h), suggesting that this step may be rate limiting for E1E2 oligomerization. Both HCV glycoproteins associated rapidly with calnexin, but dissociation was slow, consistent with the slow folding and assembly of E1E2 glycoprotein complexes. These results suggest a role for prolonged association with calnexin in the folding and assembly of HCV glycoprotein heterodimer complexes.
Dubuisson
J
J
Unité d'Oncologie Moléculaire, Centre National de la Recherche Scientifique--URA-1160, Institut Pasteur de Lille, France.
Rice
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D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D037281:Calnexin; D002460:Cell Line; D004220:Disulfides / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D017510:Protein Folding; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000378:metabolism
D002135:Calcium-Binding Proteins; D004220:Disulfides; C087035:E1 protein, Hepatitis C virus; D006023:Glycoproteins; D018832:Molecular Chaperones; C097010:NS2 protein, Hepatitis C virus; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; D037281:Calnexin; C091015:glycoprotein E2, Hepatitis C virus