Gretch D R, Kari B, Rasmussen L, Gehrz R C, Stinski M F
Department of Microbiology, University of Iowa, Iowa City 52242.
J Virol. 1988 Mar;62(3):875-81. doi: 10.1128/JVI.62.3.875-881.1988.
Several disulfide-linked glycoprotein complexes were identified in the envelope of human cytomegalovirus (HCMV). These glycoprotein complexes were fractionated by rate-zonal centrifugation in sucrose density gradients in the presence of detergents. Fractionated glycoproteins and complexes were immunoprecipitated with three different monoclonal antibodies specific for HCMV glycoproteins and a rabbit polyclonal antiserum prepared against detergent-extracted virion and dense-body envelope glycoproteins. Three distinct families of disulfide-linked glycoprotein complexes were observed and designated glycoprotein complex gcI, gcII, and gcIII. The gcI family, recognized by monoclonal antibody 41C2 under nonreducing conditions, consisted of three complexes with approximate molecular masses of 250 to 300, 190, and 160 kilodaltons (kDa). These complexes consistently sediment more rapidly than other HCMV glycoproteins or complexes in sucrose density gradients. Upon reduction of the gcI family, two size classes of glycoproteins with average molecular masses of 93 to 130 and 55 kDa were observed. The gcII family was recognized by monoclonal antibody 9E10. Under nonreducing conditions, as many as six electrophoretic forms were observed for gcII. When reduced, the major component of the gcII family was a heterogeneous glycoprotein designated gp47-52. The gcIII family was recognized by monoclonal antibody 1G6. It consisted of a complex of approximately 240 kDa without reduction of disulfide bonds. When reduced, two glycoprotein size classes with average molecular masses of 145 and 86 kDa were observed. Polyclonal antiserum R-7 reacted strongly with the gcI and gcIII families, but weakly with the gcII family.
在人巨细胞病毒(HCMV)包膜中鉴定出了几种二硫键连接的糖蛋白复合物。在去污剂存在的情况下,通过蔗糖密度梯度中的速率区带离心对这些糖蛋白复合物进行分级分离。分级分离的糖蛋白和复合物用三种针对HCMV糖蛋白的不同单克隆抗体以及针对去污剂提取的病毒体和致密体包膜糖蛋白制备的兔多克隆抗血清进行免疫沉淀。观察到三个不同的二硫键连接的糖蛋白复合物家族,分别命名为糖蛋白复合物gcI、gcII和gcIII。在非还原条件下,被单克隆抗体41C2识别的gcI家族由三种复合物组成,其近似分子量分别为250至300、190和160千道尔顿(kDa)。在蔗糖密度梯度中,这些复合物始终比其他HCMV糖蛋白或复合物沉降得更快。对gcI家族进行还原后,观察到两种分子量平均为93至130和55 kDa的糖蛋白大小类别。gcII家族被单克隆抗体9E10识别。在非还原条件下,观察到gcII多达六种电泳形式。还原后,gcII家族的主要成分是一种异质性糖蛋白,命名为gp47 - 52。gcIII家族被单克隆抗体1G6识别。它由一个约240 kDa的复合物组成,二硫键未还原。还原后,观察到两种平均分子量为145和86 kDa的糖蛋白大小类别。多克隆抗血清R - 7与gcI和gcIII家族反应强烈,但与gcII家族反应较弱。