SYNTHESIS AND PROCESSING OF GLYCOPROTEINS OF VARICELLA-ZOSTER VIRUS (VZV) AS STUDIED WITH MONOCLONAL-ANTIBODIES TO VZV ANTIGENS
SYNTHESIS AND PROCESSING OF GLYCOPROTEINS OF VARICELLA-ZOSTER VIRUS (VZV) AS STUDIED WITH MONOCLONAL-ANTIBODIES TO VZV ANTIGENS
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DOI：
10.1016/0042-6822(83)90175-7
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发表时间：
1983-01-01
期刊：
影响因子：
3.7
通讯作者：
TAKAHASHI, M
中科院分区：
文献类型：
--
作者：
OKUNO, T;YAMANISHI, K;TAKAHASHI, M
水痘带状疱疹病毒（VZV）含有4种主要糖蛋白，分别命名为gp1（115K）、gp2（100 - 80K）、gp3（64K）和gp5（55K）。目前的研究集中在这些糖蛋白的合成和加工使用单克隆抗体。已经建立了27株分泌抗VZV蛋白单克隆抗体的小鼠杂交瘤。用[35S]甲硫氨酸、[3H]葡糖胺、[125I]标记的病毒抗原进行放射免疫沉淀，然后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），进一步表征抗体。这些杂交瘤的12个克隆产生抗体的VZV的糖基化多肽，可以分为3组的电泳图谱的基础上。第一组与感染的人胚成纤维细胞、感染细胞的表面膜和病毒体上的表观分子量为94K和83K的多肽（推测两者对应于gp 2）特异性反应。第2组在感染细胞中沉淀出分子量为94K、83K、55K（对应于gp5）的多肽，在感染细胞表面膜上沉淀出分子量为94K、83K、55K、45K的多肽，在病毒粒子上沉淀出分子量为94K、83K、55K的多肽。第3组与感染细胞中分子量为116K、106K和64K的多肽（对应于gp3）反应，与感染细胞表面膜和病毒粒子上的64K多肽反应。通过脉冲追踪实验，前两组抗体分别沉淀出分子量为75K和49K的新多肽，推测为前体蛋白。这些数据表明，gp 2，gp 3和gp 5的前体蛋白在感染细胞中合成，糖基化，并且产物蛋白在感染细胞的表面膜以及病毒体上表达。在感染细胞的培养液中检测到一种分子量为45K的糖蛋白，它是由分子量为55K的多肽（gp 5）衍生而来的。
Varicella-zoster virus (VZV) contains 4 major glycoproteins designated as gp 1 (115K), gp 2 (100-80K), gp 3 (64K) and gp 5 (55K). The present studies focused on the synthesis and processing of these glycoproteins using monoclonal antibodies. Twenty-seven mouse hybridomas secreting monoclonal antibodies against VZV proteins have been established. The antibodies were characterized further by radioimmunoprecipitation with [35S]methionine, [3H]glucosamine, [125I]labeled viral antigens followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Twelve clones of these hybridomas produced antibodies to glycosylated polypeptides of VZV, which could be classified into 3 groups on the basis of the electrophoretic patterns. The 1st group reacted specifically with polypeptides with apparent MW of 94K and 83K (both presumed to correspond to gp 2) in infected human embryo fibroblast cells, on the surface membrane of infected cells and on the virions. The 2nd group precipitated polypeptides with MW of 94K, 83K, 55K (corresponding to gp 5) in infected cells, and 94K, 83K, 55K, 45K on the surface membrane of infected cells and 94K, 83K and 55K on the virions. The 3rd group reacted with polypeptides with MW of 116K, 106K and 64K (corresponding to gp 3) in infected cells, 64K on the surface membrane of infected cells and on the virions. By pulse-chase experiments, antibodies from the former 2 groups precipitated new polypeptides with MW of 75K and 49K, respectively, which were presumed to be precursor proteins. These data suggest that the precursor proteins of gp 2, gp 3 and gp5 are synthesized in infected cells, glycosylated and the product proteins are expressed on the surface membrane of infected cells as well as on the virions. It was also found that a glycoprotein of 45K detected in culture fluid of infected cells was derived from the polypeptide with MW of 55K (gp 5).