Signal peptide replacements enhance expression and secretion of hepatitis C virus envelope glycoproteins

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 El/E2 expression and secretion is critical for the characterization of these glycoproteins and thus for subunit vaccine development....

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Bibliographic Details
Published in:Acta biochimica et biophysica Sinica 2011-02, Vol.43 (2), p.96-102
Main Authors: Wen, Bo, Deng, Yao, Guan, Jie, Yan, Weizheng, Wang, Yue, Tan, Wenjie, Gao, Jimin
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Cell Line
Gene Expression
Glycoproteins - genetics
Glycoproteins - metabolism
Glycoproteins - secretion
Hepacivirus - metabolism
Humans
Molecular Sequence Data
Protein Sorting Signals - genetics
Viral Envelope Proteins - genetics
Viral Envelope Proteins - metabolism
Viral Envelope Proteins - secretion
Western印迹
丙型肝炎病毒
信号肽序列
包膜糖蛋白
哺乳动物细胞
组织纤溶酶原激活物
雪花莲凝集素
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Summary: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 El/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.
ISSN:1672-9145
1745-7270
DOI:10.1093/abbs/gmq117