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登革2型病毒感染的非洲绿猴肾细胞和白纹伊蚊细胞中蛋白质和糖蛋白的合成

Synthesis of proteins and glycoproteins in dengue type 2 virus-infected vero and Aedes albopictus cells.

作者信息

Smith G W, Wright P J

出版信息

J Gen Virol. 1985 Mar;66 ( Pt 3):559-71. doi: 10.1099/0022-1317-66-3-559.

Abstract

Fifteen proteins were detected in Vero cells infected by dengue type 2 (DEN-2) virus that were not observed in mock-infected cells, namely P98, p82, P67, GP60, gp54, GP46, p30, p28, gp22, GP20, p18, gp16, p15, p14 and gp13. With the exceptions of gp54 and gp13, polypeptides corresponding to those listed above were also observed in DEN-2 virus-infected Aedes albopictus C6/36 cells. Pulse-chase labelling experiments suggested a possible precursor-product relationship between p30 and p28, and between gp22 and GP20. Peptide mapping and immunoprecipitation experiments showed that the major glycoproteins GP60, GP46 and GP20 were unrelated. Immunoprecipitations of infected cells with antiserum prepared against the DEN-2 soluble complement-fixing (SCF) antigen demonstrated that this antigen is equivalent to the non-structural glycoprotein GP46. The envelope glycoprotein (E) from virus grown in C6/36 cells migrated faster through polyacrylamide gels containing SDS than E from virus grown in Vero cells. [3H]Mannose-labelled glycopeptides of GP60, GP46 and GP20 were separated by gel filtration and by electrophoresis in Tris-borate gels; in addition, the polypeptides synthesized in infected cells in the presence of tunicamycin were analysed. The results revealed heterogeneity among the glycan units of GP60 and GP46.

摘要

在感染2型登革热病毒(DEN-2)的Vero细胞中检测到15种在模拟感染细胞中未观察到的蛋白质,即P98、p82、P67、GP60、gp54、GP46、p30、p28、gp22、GP20、p18、gp16、p15、p14和gp13。除了gp54和gp13之外,在感染DEN-2病毒的白纹伊蚊C6/36细胞中也观察到了与上述蛋白质相对应的多肽。脉冲追踪标记实验表明p30和p28之间以及gp22和GP20之间可能存在前体-产物关系。肽图谱和免疫沉淀实验表明主要糖蛋白GP60、GP46和GP20之间没有关系。用针对DEN-2可溶性补体结合(SCF)抗原制备的抗血清对感染细胞进行免疫沉淀,结果表明该抗原等同于非结构糖蛋白GP46。在C6/36细胞中培养的病毒的包膜糖蛋白(E)在含有SDS的聚丙烯酰胺凝胶中迁移速度比在Vero细胞中培养的病毒的E更快。通过凝胶过滤和在Tris-硼酸盐凝胶中进行电泳分离了GP60、GP46和GP20的[3H]甘露糖标记的糖肽;此外,还分析了在衣霉素存在下感染细胞中合成的多肽。结果揭示了GP60和GP46聚糖单元之间的异质性。

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