Mallon V, Holowczak J A
Virology. 1985 Mar;141(2):201-20. doi: 10.1016/0042-6822(85)90252-1.
SDS-polyacrylamide gel electrophoretic analysis of plasma membranes prepared from L cells infected with radioiodinated vaccinia virus particles showed that at 2.0 hr postinfection, 125I-labeled virion polypeptides with molecular weights of 58K-60K, 32K-34K, 17K, and 12K-14K were associated with infected cell plasma membranes. By 4 hr postinfection, only the 32- to 34-kDa polypeptide, derived from infecting virus particles, could be detected on infected cell surfaces. A variety of techniques were applied to analyzing purified plasma cell membranes to define the viral antigens expressed on cell surfaces after infection, including (a) surface radioiodination of infected cells; (b) immune or Western blotting; (c) specific immunoprecipitation of viral proteins present in nonionic detergent extracts of membranes purified from [35S]methionine-labeled, virus-infected cells. It was determined that vaccinia virus-specified polypeptides with molecular weights of 78K-82K, 65K, 50K, 42K-45K, 35K-37K, 32K-34K, 30K, 20K, and 17K-18K were expressed by 3 hr postadsorption, on the plasma membranes of infected cells and were accessible to binding by exogenous antiviral antibodies. Viral antigens with molecular weights similar to those expressed on cell surfaces were secreted or shed from infected cells and could be detected in the medium harvested from virus-infected mouse L-cell cultures.
对用放射性碘化痘苗病毒颗粒感染的L细胞制备的质膜进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明,在感染后2.0小时,分子量为58K - 60K、32K - 34K、17K和12K - 14K的125I标记病毒粒子多肽与感染细胞的质膜相关。到感染后4小时,在感染细胞表面仅能检测到源自感染病毒颗粒的32至34 kDa多肽。应用了多种技术来分析纯化的质膜,以确定感染后细胞表面表达的病毒抗原,包括:(a) 感染细胞的表面放射性碘化;(b) 免疫印迹或蛋白质印迹;(c) 对从[35S]甲硫氨酸标记的、病毒感染细胞中纯化的膜的非离子去污剂提取物中存在的病毒蛋白进行特异性免疫沉淀。已确定,痘苗病毒指定的分子量为78K - 82K、65K、50K、42K - 45K、35K - 37K、32K - 34K、30K、20K和17K - 18K的多肽在吸附后3小时在感染细胞的质膜上表达,并可被外源性抗病毒抗体结合。分子量与细胞表面表达的类似的病毒抗原从感染细胞中分泌或脱落,并可在从病毒感染的小鼠L细胞培养物中收获的培养基中检测到。