Broughan J H, Wunner W H
Wistar Institute, Philadelphia, Pennsylvania, USA.
Arch Virol. 1995;140(1):75-93. doi: 10.1007/BF01309725.
Prior studies established the specificity of rabies virus receptors on BHK-21 cells based on the saturability of the receptors and on competitive binding. In the present study, we used protease-treated cells to identify the involvement of protein in the specific binding of rabies virus to these cells. In addition, biochemical characterization of n-octylglucoside solubilized BHK-21 plasma membranes demonstrated the involvement of a protease sensitive, heat insensitive, integral membrane protein or protein complex in rabies virus binding to these cells. The membrane component that binds rabies virus is associated with a high molecular weight fraction of the n-octylglucoside-plasma membrane extract isolated by gel filtration. This high molecular weight fraction (approximately 450 KDa) is enriched with a cell surface integral membrane component that comigrates with denatured bovine serum fibronectin (220 KDa). This cellular component did not bind polyclonal antisera to fibronectin in Western blot (native or denatured) or immunoprecipitation experiments. Direct and specific virus binding to high molecular weight plasma membrane protein(s) separated on Western blots further confirmed the role of a protein receptor in rabies virus binding to these cells.
先前的研究基于受体的饱和性和竞争性结合确定了狂犬病病毒受体在BHK-21细胞上的特异性。在本研究中，我们使用蛋白酶处理的细胞来确定蛋白质在狂犬病病毒与这些细胞的特异性结合中的作用。此外，对正辛基葡萄糖苷增溶的BHK-21质膜的生化特性分析表明，一种对蛋白酶敏感、对热不敏感的整合膜蛋白或蛋白复合物参与了狂犬病病毒与这些细胞的结合。与狂犬病病毒结合的膜成分与通过凝胶过滤分离的正辛基葡萄糖苷-质膜提取物的高分子量组分相关。这个高分子量组分（约450 kDa）富含一种细胞表面整合膜成分，它与变性牛血清纤连蛋白（220 kDa）一起迁移。在蛋白质印迹法（天然或变性）或免疫沉淀实验中，这种细胞成分不与纤连蛋白的多克隆抗血清结合。狂犬病病毒与蛋白质印迹法分离的高分子量质膜蛋白的直接和特异性结合进一步证实了蛋白质受体在狂犬病病毒与这些细胞结合中的作用。
