Characterization of two different human cytomegalovirus glycoproteins which are targets for virus neutralizing antibody.
Characterization of two different human cytomegalovirus glycoproteins which are targets for virus neutralizing antibody.
复制标题
两种不同的人巨细胞病毒糖蛋白的表征，它们是病毒中和抗体的靶标。
DOI：
10.1016/0042-6822(88)90271-1
复制
发表时间：
1988
期刊：
影响因子：
3.7
通讯作者：
MeriganJr,TC
中科院分区：
文献类型：
--
作者：
Rasmussen,L;Nelson,M;Neff,M;MeriganJr,TC
在以前的研究中，我们已经确定了两个病毒多肽检测的鼠单克隆抗体，中和人巨细胞病毒（CMV）AD 169的感染性。一种是86，000 Da多肽（p86），第二种是130，000和55，000 Da两种主要免疫共沉淀多肽（p130/55）的复合物。在这项研究中，我们已经表明，这两种病毒多肽是免疫无关的，并有不同的肽切割模式。我们已经将这些多肽表征为糖蛋白，并研究了它们在人胚肺细胞中的生物合成。在p86和pl 30/55上发现的寡糖通过内切糖苷酶消化表征为N-连接的高甘露糖碳水化合物。糖基化抑制剂用于进一步表征寡糖。衣霉素抑制内质网上N-连接寡糖的生物合成，抑制p86和p130/55的感染性和生物合成。只有在L-[35 S]蛋氨酸脉冲标记的条件下，才能检测到衣霉素处理的培养物中的低糖基化形式。莫能菌素抑制高尔基体中糖蛋白从简单形式到复杂形式的修饰，在对病毒蛋白合成没有影响的浓度下降低病毒感染性，但不改变p86或p130/55的表观分子量。在免疫印迹中，寡糖对p86的体外免疫反应性至关重要。然而，内切糖苷酶F处理的p86在豚鼠中诱导病毒中和抗体方面与天然形式相当。内切糖苷酶F处理的p130/55在Western印迹中保留了其结合抗体的能力。
In previous studies we have identified two viral polypeptides detected by murine monoclonal antibodies which neutralize the infectivity of human cytomegalovirus (CMV) AD169. One is an 86,000-Da polypeptide (p86) and the second is a complex of two major coimmunoprecipitating polypeptides of 130,000 and 55,000 Da (p130/55). In this study we have shown that the two viral polypeptides are immunologically unrelated and have distinct peptide cleavage patterns. We have characterized these polypeptides as glycoproteins and studied their biosynthesis in human embryonic lung cells. The oligosaccharides found on both the p86 and the pl30/55 were characterized by endoglycosidase digestion as N-linked high-mannose carbohydrates. Inhibitors of glycosylation were used to further characterize the oligosaccharides. Tunicamycin, which inhibits the biosynthesis of N-linked oligosaccharides on the endoplasmic reticulum, inhibited both the infectivity and biosynthesis of the p86 and p130/55. The underglycosylated forms in tunicamycin-treated cultures could be detected only under conditions of pulse-labeling with L-[35S)methionine. Monensin, which inhibits the modification of glycoproteins from simple to complex forms in the Golgi, reduced viral infectivity at concentrations which had no effect on viral protein synthesis, but did not alter the apparent molecular weight of either the p86 or the p130/55. The oligosaccharides were critical for thein vitroimmunologic reactivity of the p86 in immunoblots. However, endoglycosidase F-treated p86 was comparable to the native form in inducing virus neutralizing antibody in guinea pigs. Endoglycosidase F-treated p130/55 retained its ability to bind antibody in Western blots.