Division of Biochemical Virology and Membrane Research Department of Ophthalmology University of Pennsylvania School of Medicine Scheie Eye Institute Philadelphia, Pennsylvania 19104 USA
Abstract:
The envelope membrane glycoprotein gC of HSV-1 was purified from Triton X-100 extracts of virus-infected BHK-21 or HEp-2 cells by a single step immuno-affinity column using monoclonal anti-gC antibody. The analysis of the purified 3H]G1cN labeled glycoprotein gC (by gel filtration on Bio-Gel P4) before and after digestion with endo-β-N-acetylglucosaminidase (endo D) indicated that gC contains Asn-linked “complex type” oligosaccharides. No “high mannose” type oligosaccharides were detected. Fractionation of radio-labeled glycopeptides of gC on a column of concanavalin A-sepharose suggested that glycopeptides have “diantennary” and “triantennary” and/or “tetra antennary” structures. Tunicamycin inhibited the incorporation of 14C]GalN or 3H]GlcN into gC in HSV-1 infected BHK-21 or HEp-2 cells. Gel filtration analysis of 3H]GlcN labeled gC following β-elimination reaction failed to indicate O-glycosidically linked oligosaccharides.