Herold B C, WuDunn D, Soltys N, Spear P G
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois 60611.
J Virol. 1991 Mar;65(3):1090-8. doi: 10.1128/JVI.65.3.1090-1098.1991.
The purpose of this study was to identify the herpes simplex virus glycoprotein(s) that mediates the adsorption of virions to cells. Because heparan sulfate moieties of cell surface proteoglycans serve as the receptors for herpes simplex virus adsorption, we tested whether any of the viral glycoproteins could bind to heparin-Sepharose in affinity chromatography experiments. Two glycoproteins, gB and gC, bound to heparin-Sepharose and could be eluted with soluble heparin. In order to determine whether virions devoid of gC or gB were impaired for adsorption, we quantitated the binding of wild-type and mutant virions to cells. We found that at equivalent input concentrations of purified virions, significantly fewer gC-negative virions bound to cells than did wild-type or gB-negative virions. In addition, the gC-negative virions that bound to cells showed a significant delay in penetration compared with wild-type virus. The impairments in adsorption and penetration of the gC-negative virions can account for their reduced PFU/particle ratios, which were found to be about 5 to 10% that of wild-type virions, depending on the host cell. Although gC is dispensable for replication of herpes simplex virus in cell culture, it clearly facilitates virion adsorption and enhances infectivity by about a factor of 10.
本研究的目的是鉴定介导病毒粒子吸附到细胞上的单纯疱疹病毒糖蛋白。由于细胞表面蛋白聚糖的硫酸乙酰肝素部分充当单纯疱疹病毒吸附的受体，我们在亲和色谱实验中测试了是否有任何病毒糖蛋白能与肝素-琼脂糖结合。两种糖蛋白gB和gC与肝素-琼脂糖结合，并能用可溶性肝素洗脱。为了确定缺乏gC或gB的病毒粒子在吸附方面是否受损，我们对野生型和突变型病毒粒子与细胞的结合进行了定量。我们发现，在纯化病毒粒子的输入浓度相等时，与野生型或gB阴性病毒粒子相比，gC阴性病毒粒子与细胞结合的数量明显更少。此外，与野生型病毒相比，结合到细胞上的gC阴性病毒粒子在穿透方面有显著延迟。gC阴性病毒粒子在吸附和穿透方面的损伤可以解释其降低的空斑形成单位/粒子比率，根据宿主细胞的不同，该比率约为野生型病毒粒子的5%至10%。虽然gC对于单纯疱疹病毒在细胞培养中的复制不是必需的，但它显然促进了病毒粒子的吸附，并将感染性提高了约10倍。
