Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells.
Truncated and full-length versions of the hepatitis C virus protein domain encoding a presumptive envelope glycoprotein designated E2/NS1 were stably expressed in CHO cell lines. Characterization of the processing events involved in the maturation of E2/NS1 revealed that a high-mannose form resident in the endoplasmic reticulum was the most abundant form detected intracellularly. The ionophore carboxyl cyanide m-chlorophenyl-hydrazone was used to show that the E2/NS1 glycoprotein resided in the endoplasmic reticulum. The full-length form of E2/NS1 appeared to be cell-associated and could not be detected as a secreted product. C-terminal truncated molecules could be detected in the extracellular media as fully processed glycoproteins containing terminal sialic acid additions. These truncated glycoproteins are predicted to be biologically relevant targets of the host immune response and are therefore potential subunit vaccine candidates.
Spaete
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RR
Chiron Corporation, Emeryville, California 94608-2916.
Alexander
D
D
Rugroden
M E
ME
Choo
Q L
QL
Berger
K
K
Crawford
K
K
Kuo
C
C
Leng
S
S
Lee
C
C
Ralston
R
R
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D001212:Asialoglycoproteins / Q000378:metabolism; D001483:Base Sequence; D016466:CHO Cells; D002213:Capsid / Q000235:genetics* / Q000378:metabolism; D002451:Cell Compartmentation; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D016174:Hepacivirus / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D017361:Viral Nonstructural Proteins
D000956:Antigens, Viral; D001212:Asialoglycoproteins; D009838:Oligodeoxyribonucleotides; D011506:Proteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins