Hobman T C, Qiu Z Y, Chaye H, Gillam S
Department of Pathology, University of British Columbia Research Center, Vancouver, Canada.
Virology. 1991 Apr;181(2):768-72. doi: 10.1016/0042-6822(91)90915-x.
cDNA clones encoding the envelope glycoprotein E1 of rubella virus (RV) were altered by site-directed mutagenesis at consensus sites for addition of N-linked glycans. The resulting plasmids were introduced into COS cells and the mutant E1 proteins were analyzed by indirect immunofluorescence, radioimmunoprecipitation, and immunoblotting. We found that RV E1 contains three N-linked oligosaccharides, each approximately 2 kDa in size. Although lack of glycosylation did not appear to affect targeting of E1 to the Golgi region, mutants lacking N-linked glycans at Asn 177 and Asn 209 failed to bind anti-E1 antibodies under nonreducing conditions. Our results suggest that glycosylation may be important for expression of important immunologic epitopes on RV E1.
通过定点诱变，在风疹病毒（RV）包膜糖蛋白E1编码cDNA克隆的N-连接聚糖添加共有位点进行改造。将所得质粒导入COS细胞，并通过间接免疫荧光、放射免疫沉淀和免疫印迹分析突变型E1蛋白。我们发现RV E1含有三个N-连接寡糖，每个大小约为2 kDa。虽然糖基化缺失似乎不影响E1靶向高尔基体区域，但在Asn 177和Asn 209处缺乏N-连接聚糖的突变体在非还原条件下无法结合抗E1抗体。我们的结果表明，糖基化对于RV E1上重要免疫表位的表达可能很重要。
