title: A structural investigation of the Epstein-Barr (EB) virus membrane antigen glycoprotein, gp340.
abstract: Epstein-Barr (EB) virus membrane antigen (MA) glycoprotein (gp340) purified by a molecular weight-based technique has been subjected to biochemical analysis. Following treatment with glycosidases or tunicamycin during synthesis, the carbohydrate moiety was found to be made up of both O-linked and N-linked types and to constitute about 50% of the molecular mass. Digestion studies with neuraminidase and oligosaccharidase have indicated that the molecule is heavily sialated with most of the sialic acid located on the O-linked sugars. The high carbohydrate content of gp340 appears to confer resistance to proteolysis; thus, V8 protease was only effective at concentrations above 1 mg/ml when three large fragments of mol. wt. 330K, 190K and 160K were generated. Removal of sialic acid before V8 protease digestion did not alter this pattern nor affect the antigenicity of the digestion fragments. Antigenicity of the intact molecule was likewise unaffected by removal of sialic acid nor were the O-linked and N-linked carbohydrate moieties essential for this property. The binding of virus-neutralizing human sera and monoclonal antibody by gp340 from which either O-linked or N-linked sugars had been removed seems to indicate that the sites on the molecule that generate the neutralizing antibodies are present in the protein component. The significance of these results is discussed in relation to the development of a subunit vaccine against EB virus.
mesh_terms: D000818:Animals; D002143:Callitrichinae; D002460:Cell Line; D006026:Glycoside Hydrolases; D004854:Herpesvirus 4, Human / Q000187:drug effects / Q000276:immunology*; D008214:Lymphocytes; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D009844:Oligosaccharides; D014759:Viral Envelope Proteins; D014415:Tunicamycin; D006026:Glycoside Hydrolases