Isolation and characterization of a human cytomegalovirus glycoprotein containing a high content of O-linked oligosaccharides.
Several disulfide linked glycoprotein complexes were extracted from human cytomegalovirus with a non-ionic detergent and separated by anion exchange high performance liquid chromatography (HPLC). One complex had a molecular weight of 93,000 and was classified as gCII-93. Another complex had a molecular weight greater than 200,000 and was classified as gCII-200. Both complexes immunoprecipitated with a monoclonal antibody (9E10). A third set of complexes (classified as gC-I) immunoprecipitated with another monoclonal antibody (41C2). Isolated complexes were reduced, alkylated, and individual glycoproteins separated by gel-filtration HPLC. Glycoproteins with molecular weights of 50-52,000 from gCII-93 and gCII-200 appeared to be the same glycoprotein since they could be immunoprecipitated by 9 E 10 and had identical peptide maps. The amino sugar content of these glycoproteins was compared to that of higher molecular weight glycoproteins obtained from gCII-200 and to a glycoprotein with a molecular weight of 93,000 (gp93 (I] from gCI. Glycoproteins with molecular weights of 50-52,000 from gCII-93 and gCII-200 contained similar amounts of galactosamine (GalN), glucosamine (GlcN) and sialic acid. However, they contained 2-3 times more GalN than any other glycoprotein from gCII-200 and 10 times more GalN than was detected in gp93 (I). All glycoproteins from gCII-93 or gCII-200 also contained more sialic acid when compared to gp93 (I). GalN in these glycoproteins was present in O-linked oligosaccharides. This was demonstrated by release of low molecular weight oligosaccharides from high molecular weight glycopeptides by mild base hydrolysis and the conversion of GalN to galactosaminitol. Thus, gp52(II) appears to have a unique phenotype marked by a high amount of O-linked oligosaccharides.
Kari
B
B
Biomedical Research Center, Children's Hospital, St. Paul, Minnesota.
Gehrz
R
R
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D003587:Cytomegalovirus / Q000032:analysis* / Q000276:immunology; D005688:Galactosamine / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000032:analysis*; D010449:Peptide Mapping; D012794:Sialic Acids / Q000032:analysis; D014764:Viral Proteins / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D009844:Oligosaccharides; D012794:Sialic Acids; D014764:Viral Proteins; D005688:Galactosamine; D019158:N-Acetylneuraminic Acid; D005944:Glucosamine