SYNTHESIS AND PROCESSING OF GLYCOPROTEIN-GG OF HERPES-SIMPLEX VIRUS TYPE-2
SYNTHESIS AND PROCESSING OF GLYCOPROTEIN-GG OF HERPES-SIMPLEX VIRUS TYPE-2
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DOI：
10.1128/jvi.54.3.825-832.1985
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发表时间：
1985-01-01
影响因子：
5.4
通讯作者：
HUTTFLETCHER, LM
中科院分区：
文献类型：
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作者：
BALACHANDRAN, N;HUTTFLETCHER, LM
单克隆抗体13 α C5 -1-A11从用[35 S]甲硫氨酸或[3 H]葡糖胺标记的单纯疱疹病毒2型感染的BHK-21细胞的提取物中免疫沉淀MW为108，000和120，000的2种主要多肽。在脉冲追踪实验中，将两种标记物从120，000-MW肽（120 K肽）追踪到108 K分子中。内切糖苷酶H（endo H）将120 K肽还原为112 K肽，但不影响108 K肽。用单克隆抗体AP-1获得了类似的谱，AP-1与92 K糖蛋白gG反应，gG映射到基因组的短的独特区域。交叉吸收实验表明，这两种抗体与相同的肽反应，表明120 K肽是部分糖基化的高甘露糖型前体的gG（pgG 1）。从莫能菌素处理的细胞的免疫沉淀表明，pgG 1（120 K）可以进行肽裂解，形成74 K高甘露糖型肽（pgG 2），这74 K肽可以进一步加工成endoH-抗性110 K-116 K肽。在衣霉素存在下，gG（108 K）被110 K和105 K肽取代，所述110 K和105 K肽对内切H和内切糖苷酶F均具有抗性。在衣霉素存在下，105 K肽是唯一被[3 H]半乳糖或[3 H]葡糖胺标记的分子，并且没有肽被[3 H]甘露糖标记，表明105 K肽中可能存在O-连接的糖。这些结果表明，非糖基化前体110 K肽的共翻译糖基化导致高甘露糖型pgG 1（120 K），其可能经历肽裂解。这种假定的裂解产物然后可以通过糖的修剪和添加复合物和可能的O-连接糖而成熟为gG（108 K）;高甘露糖型pgG 2（74 K）可能是在此过程中形成的中间肽。
Monoclonal antibody 13.alpha.C5-1-A11 immunoprecipitated 2 major polypeptides of MW 108,000 and 120,000 from extracts of herpes simplex virus type 2-infected BHK-21 cells labeled with [35S]methionine or [3H]glucosamine. In pulse-chase experiments, both labels were chased from the 120,000-MW peptide (120K peptide) into the 108K molecule. Endoglycosidase H (endo H) reduced the 120K peptide to a 112K peptide but did not affect the 108K peptide. Similar profiles were obtained with monoclonal antibody AP-1 which reacts with a 92K glycoprotein, gG, which maps to the short unique regions of the genome. Cross-absorption experiments indicated that both antibodies reacted with the same peptides, suggesting that the 120K peptide is a partially glycosylated high-mannose-type precursor of gG (pgG1). Immunoprecipitation from monensin-treated cells indicated that pgG1(120K) may undergo peptide cleavage to form a 74K high-mannose-type peptide (pgG2) and that this 74K peptide may be further processed into an endoH-resistant 110K-116K peptide. In the presence of tunicamycin, gG(108K) was replaced by 110K and 105K peptides which were resistant to both endo H and endoglycosidase F. The 105K peptide was the only molecule labeled by [3H]galactose or [3H]glucosamine in the presence of tunicamycin, and none of the peptides were labeled with [3H]mannose, indicating the probable presence of O-linked sugars in the 105K peptide. These results imply that cotranslational glycosylation of the unglycosylated precursor 110K peptide results in the high-mannose-type pgG1(120K), which probably undergoes peptide cleavage. This putative cleavage product may then mature into gG(108K) by the trimming of sugars and the addition of complex and probably O-linked sugars; the high-mannose-type pgG2(74K) is probably an intermediate peptide formed in this process.