Analysis of hepatitis C virus capsid, E1, and E2/NS1 proteins expressed in insect cells.
The baculovirus/insect cell expression system was used to express the capsid protein and glycoproteins (e1 and e2/NS1) of the hepatitis C virus (HCV). Each polypeptide domain was expressed individually using two different constructs varying at their carboxy termini in order to retain or delete hydrophobic domains that may be involved in membrane association. The capsid proteins were transported to the nucleus where they formed a single large crystal-like inclusion. The capsid proteins were phosphorylated in insect cells. The e1 and e2 polypeptides were present in both the soluble and insoluble cellular fractions. Deletion of a hydrophobic domain in the carboxy terminus of e2 resulted in the polypeptide becoming soluble but not secreted. Deletion of the carboxy terminus of e1 had no effect on solubility. Both e1 and e2 were glycosylated, with variable glycosylation of e1 giving rise to a series of polypeptides varying in apparent molecular weight. Co-infection of insect cells with viruses expressing e1 and e2 resulted in a complex that permitted the coimmunoprecipitation of e1 with antibodies to e2 and vice versa. Immunofluorescence staining of insect cells expressing e1 and e2 indicated that reactivity to e2 was more prevalent in anti-HCV positive human sera.
Lanford
R E
RE
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas 78228.
Notvall
L
L
Chavez
D
D
White
R
R
Frenzel
G
G
Simonsen
C
C
Kim
J
J
D000818:Animals; D002213:Capsid / Q000032:analysis / Q000096:biosynthesis*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D009036:Moths; D014162:Transfection; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis*
D006023:Glycoproteins; D014764:Viral Proteins