Hunt L A
J Virol. 1980 Aug;35(2):362-70. doi: 10.1128/JVI.35.2.362-370.1980.
To determine the particular intracellular steps in the glycosylation of the vesicular stomatitis virus (VSV) glycoprotein that were altered in several lectin-resistant CHO cell lines, VSV-infected parental and mutant cells were pulse-labeled for 30 and 120 min with [3H]mannose and [3H]glucosamine. Cell-associated viral glycopeptides were analyzed by gel filtration combined with specific glycosidase digestions and compared with the corresponding mature virion oligosaccharides. The intracellular glycosylation of the VSV glycoprotein in a mutant cell line resistant to phytohemagglutinin was identical to that in the normal cells except for a complete block in processing at a specific step in the final trimming of the oligomannosyl core from five to three mannoses. The results demonstrated that a double-mutant cell line selected from the phytohemagglutinin-resistant cells for resistance to concanavalin A had an additional defect in one of the earliest stages of glycosylation, resulting in smaller precursor oligosaccharides linked to protein.
为了确定水泡性口炎病毒（VSV）糖蛋白糖基化过程中在几种凝集素抗性CHO细胞系中发生改变的特定细胞内步骤，用[3H]甘露糖和[3H]葡糖胺对感染VSV的亲本细胞和突变细胞进行30分钟和120分钟的脉冲标记。通过凝胶过滤结合特异性糖苷酶消化分析细胞相关的病毒糖肽，并与相应的成熟病毒体寡糖进行比较。除了在寡甘露糖核心从五个甘露糖最终修剪到三个甘露糖的特定步骤中加工完全受阻外，对植物血凝素抗性的突变细胞系中VSV糖蛋白的细胞内糖基化与正常细胞中的相同。结果表明，从对植物血凝素抗性的细胞中选择的对刀豆球蛋白A抗性的双突变细胞系在糖基化的最早阶段之一存在额外缺陷，导致与蛋白质连接的前体寡糖较小。
