Marriott S J, Griffith G R, Consigli R A
J Virol. 1987 Feb;61(2):375-82. doi: 10.1128/JVI.61.2.375-382.1987.
Polyomavirus receptor moieties were extracted from the surfaces of mouse kidney cells with the nonionic detergent octyl-beta-D-glucopyranoside. Following extraction with this detergent, mouse kidney cells were refractory to polyomavirus infection. Binding studies demonstrated that this loss of susceptibility resulted from extraction of a peripheral membrane protein or proteins required for proper virus attachment to and infection of mouse kidney cells. Infection of extracted mouse kidney cells returned following a 2-h recovery period. However, the presence of cycloheximide or tunicamycin in the recovery media interfered with recovery from infection. Cells could be infected immediately after extraction by supplying them with the extracted moieties prior to or concomitant with infection. A complex of polyomavirus and the extracted receptor protein was formed by in vitro incubation and was stable in sucrose gradient analysis. Functional receptor moieties were prepared in the form of liposomes from the detergent extract. The virus-receptor complex was immunoprecipitated with anti-polyomavirus immunoglobulin G, and the portion of the complex contributed by the cell was identified. Immunoblot analysis of the mouse kidney cell detergent extract with a receptor-specific 125I-labeled anti-idiotypic antibody or 125I-labeled polyomavirus demonstrated several reactive proteins. Attachment of polyomavirus to mouse kidney cells, followed by extraction of the virus-receptor complex, identified polyomavirus-binding proteins similar to those observed in in vitro binding. Proteins with molecular weights of approximately 95,000, 50,000 and 25,000 to 30,000 were consistently observed in all receptor assays. The relationship between these proteins and their possible involvement as the cell receptor for polyomavirus are discussed.
用非离子去污剂辛基-β-D-吡喃葡萄糖苷从小鼠肾细胞表面提取多瘤病毒受体部分。用这种去污剂提取后，小鼠肾细胞对多瘤病毒感染具有抗性。结合研究表明，这种敏感性丧失是由于提取了病毒正确附着和感染小鼠肾细胞所需的一种或多种外周膜蛋白。经过2小时的恢复期后，提取的小鼠肾细胞恢复了感染能力。然而，恢复培养基中存在放线菌酮或衣霉素会干扰感染后的恢复。在提取后立即为细胞提供提取的部分，细胞即可被感染，无论是在感染前还是感染时提供。通过体外孵育形成多瘤病毒与提取的受体蛋白的复合物，并且在蔗糖梯度分析中该复合物是稳定的。从去污剂提取物中以脂质体的形式制备功能性受体部分。用抗多瘤病毒免疫球蛋白G对病毒-受体复合物进行免疫沉淀，并鉴定复合物中由细胞贡献的部分。用受体特异性的125I标记的抗独特型抗体或125I标记的多瘤病毒对小鼠肾细胞去污剂提取物进行免疫印迹分析，结果显示有几种反应性蛋白。多瘤病毒附着于小鼠肾细胞后，提取病毒-受体复合物，鉴定出与体外结合中观察到的类似的多瘤病毒结合蛋白。在所有受体测定中始终观察到分子量约为95,000、50,000和25,000至30,000的蛋白质。讨论了这些蛋白质之间的关系以及它们作为多瘤病毒细胞受体的可能参与情况。
