Oligosaccharide chains of herpes simplex virus type 2 glycoprotein gG.2.
gG.2 glycoprotein was purified by H966 monoclonal antibodies linked to Sepharose from herpes simplex virus type 2-infected HEp-2 cells labeled with [3H] glucosamine. The glycoprotein was subjected to Pronase digestion and the glycopeptides were fractionated by Con A-Sepharose in a major fraction (88.5% of total radioactivity) unbound to the lectin gel and in a minor species which bound to the lectin as a N-linked diantennary oligosaccharide. Mild and strong acid hydrolysis of Con A-unbound and Con A-bound fractions revealed that (i) both species were highly sialylated; (ii) the Con A-unbound fraction contained mainly labeled N-acetylgalactosamine, as is the case for O-linked oligosaccharides; and (iii) the Con A-bound fraction carried the vast majority of the labeled N-acetylglucosamine present in gG.2. Three size classes of oligosaccharides were separated from mild alkaline borohydride-treated Con A-unbound glycopeptides, which accounted for about 80% of the radioactivity present in the fraction. Galactosaminitol was recovered as the major labeled product in the strong acid hydrolyzates of the oligosaccharides generated by reductive beta-elimination, indicating that they were O-glycosidically linked to the peptide backbone. Thin-layer and DEAE-Sephacel chromatography of the three O-linked oligosaccharide species indicated that disialylated tetrasaccharides and monosialylated trisaccharides were the major components, whereas neutral disaccharide was a minor component. Digestion with neuraminidase and beta-galactosidase of the O-linked oligosaccharides supported the idea that the common disaccharide core was mainly of the structure beta-galactosyl-N-acetylgalactosamine. The large occurrence of O-linked oligosaccharides differentiates this type 2-specific herpes simplex virus glycoprotein from the type-common herpesvirus glycoproteins gB, gC, and gD.
D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D005470:Fluorometry; D005944:Glucosamine / Q000378:metabolism; D006801:Humans; D008358:Mannose / Q000378:metabolism; D009844:Oligosaccharides / Q000032:analysis*; D011402:Pronase / Q000378:metabolism; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D009844:Oligosaccharides; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C038534:glycoprotein gF, herpes simplex virus type 2; D011402:Pronase; D005944:Glucosamine; D008358:Mannose