Formation of native hepatitis C virus glycoprotein complexes.
The hepatitis C virus (HCV) glycoproteins (E1 and E2) interact to form a heterodimeric complex, which has been proposed as a functional subunit of the HCV virion envelope. As examined in cell culture transient-expression assays, the formation of properly folded, noncovalently associated E1E2 complexes is a slow and inefficient process. Due to lack of appropriate immunological reagents, it has been difficult to distinguish between glycoprotein molecules that undergo productive folding and assembly from those which follow a nonproductive pathway leading to misfolding and aggregation. Here we report the isolation and characterization of a conformation-sensitive E2-reactive monoclonal antibody (H2). The H2 monoclonal antibody selectively recognizes slowly maturing E1E2 heterodimers which are noncovalently linked, protease resistant, and no longer associated with the endoplasmic reticulum chaperone calnexin. This complex probably represents the native prebudding form of the HCV glycoprotein heterodimer. Besides providing a novel reagent for basic studies on HCV virion assembly and entry, this monoclonal antibody should be useful for optimizing production and isolation of native HCV glycoprotein complexes for serodiagnostic and vaccine applications.
Deleersnyder
V
V
Unité d'oncologie moléculaire, CNRS-URA1160, Institut Pasteur de Lille, France.
Pillez
A
A
Wychowski
C
C
Blight
K
K
Xu
J
J
Hahn
Y S
YS
Rice
C M
CM
Dubuisson
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D018937:Hepatitis C Antibodies / Q000378:metabolism; D006801:Humans; D011233:Precipitin Tests; D012367:RNA, Viral / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion
D000911:Antibodies, Monoclonal; C087035:E1 protein, Hepatitis C virus; D018937:Hepatitis C Antibodies; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus