title: Signal peptide replacements enhance expression and secretion of hepatitis C virus envelope glycoproteins.
abstract: A large number of researches focused on glycoproteins E1 and E2 of hepatitis C virus (HCV) aimed at the development of anti-HCV vaccines and inhibitors. Enhancement of E1/E2 expression and secretion is critical for the characterization of these glycoproteins and thus for subunit vaccine development. In this study, we designed and synthesized three signal peptide sequences based on online programs SignalP, TargetP, and PSORT, then removed and replaced the signal peptide preceding E1/E2 by overlapping the polymerase chain reaction method. We assessed the effect of this alteration on E1/E2 expression and secretion in mammalian cells, using western blot analysis, dot blot, and Galanthus nivalis agglutinin lectin capture enzyme immunoassay. Replacing the peptides preceding E1 and E2 with the signal peptides of the tissue plasminogen activator and Gaussia luciferase resulted in maximum enhancement of E1/E2 expression and secretion of E1 in mammalian cells, without altering glycosylation. Such an advance would help to facilitate both the research of E1/E2 biology and the development of an effective HCV subunit vaccine. The strategy used in this study could be applied to the expression and production of other glycoproteins in mammalian cell line-based systems.
authors.affiliation: Wenzhou Medical College, Wenzhou, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Wen
authors.forename: Yao
authors.initials: Y
authors.lastname: Deng
authors.forename: Jie
authors.initials: J
authors.lastname: Guan
authors.forename: Weizheng
authors.initials: W
authors.lastname: Yan
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.forename: Wenjie
authors.initials: W
authors.lastname: Tan
authors.forename: Jimin
authors.initials: J
authors.lastname: Gao
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D021382:Protein Sorting Signals; D014759:Viral Envelope Proteins