Oligosaccharide Composition of Calf Rotavirus

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Abstract
The oligosaccharides of the structural glycoprotein (VP7) of calf rotavirus were characterized. The precursor of VP7 produced in infected rhesus monkey kidney MA-104 cells in the presence of tunicamycin migrated on sodium dodecyl sulfate-polyacrylamide gels with an apparent MW 6000 less than the glycosylated glycoprotein. Endoglycosidase (Endo) H digestion of the mature virus resulted in a decrease of 5000 in the MW of VP7 in 2 discrete stages. Analysis of Endo H-treated, 3H-labeled digestion products of VP7 on Bio-Gel P-4 identified an oligosaccharide of MW 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 MW 400 in the ratio of 6:1. This indicates that the oligosaccharides of VP7 consist of 4 N-linked (Man)7 residues, 2 of which occupy more exposed and 2 more cryptic positions in the VP7 molecule.