Kari B, Gehrz R
Biomedical Research Center, Children's Hospital, St. Paul, Minnesota.
Arch Virol. 1988;98(3-4):171-88. doi: 10.1007/BF01322167.
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.
用非离子型去污剂从人巨细胞病毒中提取了几种二硫键连接的糖蛋白复合物，并通过阴离子交换高效液相色谱（HPLC）进行分离。一种复合物的分子量为93,000，被归类为gCII - 93。另一种复合物的分子量大于200,000，被归类为gCII - 200。两种复合物都能用单克隆抗体（9E10）进行免疫沉淀。第三组复合物（归类为gC - I）能用另一种单克隆抗体（41C2）进行免疫沉淀。分离出的复合物经过还原、烷基化处理，然后通过凝胶过滤HPLC分离出各个糖蛋白。来自gCII - 93和gCII - 200的分子量为50 - 52,000的糖蛋白似乎是同一种糖蛋白，因为它们能被9E10免疫沉淀且具有相同的肽图。将这些糖蛋白的氨基糖含量与从gCII - 200获得的高分子量糖蛋白以及一种分子量为93,000的糖蛋白（来自gCI的gp93（I））的氨基糖含量进行了比较。来自gCII - 93和gCII - 200的分子量为50 - 52,000的糖蛋白含有相似量的半乳糖胺（GalN）、葡萄糖胺（GlcN）和唾液酸。然而，它们含有的GalN比gCII - 200的任何其他糖蛋白多2 - 3倍，比在gp93（I）中检测到的GalN多10倍。与gp93（I）相比，来自gCII - 93或gCII - 200的所有糖蛋白也含有更多的唾液酸。这些糖蛋白中的GalN存在于O - 连接的寡糖中。通过温和碱水解从高分子量糖肽中释放低分子量寡糖以及将GalN转化为半乳糖胺醇证明了这一点。因此，gp52（II）似乎具有以大量O - 连接寡糖为特征的独特表型。
