Hutchinson L, Goldsmith K, Snoddy D, Ghosh H, Graham F L, Johnson D C
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
J Virol. 1992 Sep;66(9):5603-9. doi: 10.1128/JVI.66.9.5603-5609.1992.
Antipeptide sera were used to identify a novel glycoprotein encoded by the UL53 gene of herpes simplex virus type 1 (HSV-1). The UL53 gene product is thought to play a central role in regulating membrane fusion because mutations giving rise to the syncytial phenotype, wherein cells are extensively fused, frequently map to this gene. A single 40-kDa protein, designated gK (the ninth HSV-1 glycoprotein to be described), was detected with antipeptide sera in cells infected with both wild-type and syncytial strains of HSV-1 which were labelled with [35S]methionine and [35S]cysteine or with [3H]glucosamine, and this protein was sensitive to treatment of cells with tunicamycin. With all other HSV glycoproteins studied to date, at least two glycosylated species, often differing substantially in electrophoretic mobility, have been observed in infected cells; thus, gK is unusual in this respect. The 40-kDa gK protein was also immunoprecipitated from cells infected with a recombinant adenovirus vector carrying the UL53 gene. Two glycosylated species of 39 and 41 kDa were produced when UL53 mRNA was translated in vitro in the presence of microsomes, and these proteins differed from gK produced in infected cells not only because they possessed different electrophoretic mobilities but also because they were unable to enter gels after being heated. In addition, a 36-kDa protein was detected in extracts from cells infected with HSV-2 with use of these sera.
抗肽血清被用于鉴定由单纯疱疹病毒1型（HSV-1）的UL53基因编码的一种新型糖蛋白。UL53基因产物被认为在调节膜融合中起核心作用，因为导致细胞广泛融合的合胞体表型的突变经常定位到该基因。在用[35S]甲硫氨酸和[35S]半胱氨酸或[3H]葡糖胺标记的感染了HSV-1野生型和合胞体菌株的细胞中，用抗肽血清检测到一种单一的40 kDa蛋白，命名为gK（待描述的第九种HSV-1糖蛋白），并且该蛋白对衣霉素处理细胞敏感。对于迄今研究的所有其他HSV糖蛋白，在感染细胞中观察到至少两种糖基化形式，其电泳迁移率通常有很大差异；因此，gK在这方面是不同寻常的。40 kDa的gK蛋白也从感染了携带UL53基因的重组腺病毒载体的细胞中免疫沉淀出来。当UL53 mRNA在微粒体存在下体外翻译时，产生了两种糖基化形式，分别为39 kDa和41 kDa，这些蛋白与感染细胞中产生的gK不同，不仅因为它们具有不同的电泳迁移率，还因为它们在加热后无法进入凝胶。此外，使用这些血清在感染HSV-2的细胞提取物中检测到一种36 kDa的蛋白。
