Sialylated oligosaccharides O-glycosidically linked to glycoprotein C from herpes simplex virus type 1.
Glycoprotein C (gC) was purified by immunoabsorbent from herpes simplex virus type-1-infected BHK cells labeled with [14C]glucosamine for 11 h and chased for 3 h. Glycopeptides obtained by pronase digestion of gC were fractionated by Bio-Gel filtration and concanavalin A-Sepharose chromatography. Each glycopeptide fraction was analyzed for amino sugar composition by thin-layer chromatography. The majority of radioactivity was recovered as N-acetylglucosamine, but a significant amount of labeled N-acetylgalactosamine was detected and recovered preferentially in some glycopeptide species. Mild alkaline borohydride treatment of the glycopeptides resulted in the release of small degradation products which contained N-acetylgalactosaminitol as the major labeled component and a drastic reduction of N-acetylgalactosamine in the residual glycopeptides. These results demonstrated that gC carries O-glycosidically linked oligosaccharides in addition to the N-linked di- and triantennary glycans previously described (F. Serafini-Cessi, F. Dall'Olio, L. Pereira, and G. Campadelli-Fiume, J. Virol. 51:838-844, 1984). Chromatographic behavior on DEAE-Sephacel chromatography and neuraminidase digestion of O-linked oligosaccharides indicated the presence of two major sialylated species carrying one and two sialic acid residues, respectively. The characterization of a peculiar glycopeptide species supported the notion that some of the O-linked oligosaccharides are bound to a cluster of hydroxyamino acids located near an N-glycosylation site which carries one N-linked diantennary oligosaccharide.
Dall'Olio
F
F
Malagolini
N
N
Speziali
V
V
Campadelli-Fiume
G
G
Serafini-Cessi
F
F
D003208:Concanavalin A; D006020:Glycopeptides / Q000032:analysis; D010084:Oxidation-Reduction; D011402:Pronase; D011951:Receptors, Complement / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis*; D018139:Simplexvirus / Q000032:analysis*; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000032:analysis*
D006020:Glycopeptides; D011951:Receptors, Complement; D012795:Sialoglycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; D003208:Concanavalin A; D011402:Pronase