Kouvelos K, Petric M, Middleton P J
J Gen Virol. 1984 Jul;65 ( Pt 7):1159-64. doi: 10.1099/0022-1317-65-7-1159.
The oligosaccharides of the structural glycoprotein (VP7) of calf rotavirus were characterized. The precursor of VP7 produced in infected cells in the presence of tunicamycin migrated on SDS-polyacrylamide gels with an apparent molecular weight 6000 less than the glycosylated glycoprotein. Endoglycosidase (Endo) H digestion of the mature virus resulted in a decrease of 5000 in the molecular weight of VP7 in two discrete stages. Analysis of Endo H-treated, 3H-labelled digestion products of VP7 on Bio-Gel P-4 identified an oligosaccharide of molecular weight 1350 as the predominant form. Further treatment of the digest with mannosidase and analysis on Bio-Gel P-2 columns indicated that the oligosaccharide was digested into a free mannose and an oligosaccharide of molecular weight 400 in the ratio of 6:1. This indicates that the oligosaccharides of VP7 consist of four N-linked (Man)7 residues, two of which occupy more exposed and two more cryptic positions in the VP7 molecule.
对牛轮状病毒结构糖蛋白(VP7)的寡糖进行了表征。在衣霉素存在的情况下,感染细胞中产生的VP7前体在SDS-聚丙烯酰胺凝胶上迁移时,其表观分子量比糖基化糖蛋白小6000。成熟病毒经内切糖苷酶(Endo)H消化后,VP7的分子量分两个离散阶段降低了5000。在Bio-Gel P-4上对经Endo H处理的、3H标记的VP7消化产物进行分析,确定分子量为1350的寡糖为主要形式。用甘露糖苷酶对消化产物进一步处理,并在Bio-Gel P-2柱上进行分析,结果表明该寡糖以6:1的比例被消化成游离甘露糖和分子量为400的寡糖。这表明VP7的寡糖由四个N-连接的(Man)7残基组成,其中两个占据VP7分子中更暴露的位置,另外两个占据更隐蔽的位置。