Characterization of protein involvement in rabies virus binding to BHK-21 cells.
Characterization of protein involvement in rabies virus binding to BHK-21 cells.
复制标题
狂犬病病毒与 BHK-21 细胞结合所涉及的蛋白质特征。
DOI：
10.1007/bf01309725
复制
发表时间：
1995
影响因子：
2.7
通讯作者：
Wunner,WH
中科院分区：
文献类型：
--
作者：
Broughan,JH;Wunner,WH
以前的研究证实了狂犬病病毒受体在BHK-21细胞上的特异性是基于受体的饱和性和竞争结合。在目前的研究中，我们使用蛋白酶处理的细胞来鉴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，N-羧基葡萄糖苷溶解的BHK-21质膜的生化特性表明，狂犬病病毒与这些细胞结合时，参与了一种酶敏感的、热不敏感的完整膜蛋白或蛋白复合体的参与。结合狂犬病病毒的膜成分与通过凝胶过滤分离的正辛基葡萄糖苷-质膜提取物的高相对分子质量部分有关。这种高分子量组分(∼450KDa)富含细胞表面完整的膜成分，它与变性的牛血清纤维连接蛋白(220KDa)迁移。在Western印迹(天然的或变性的)或免疫沉淀实验中，这种细胞成分不能将多克隆抗血清与纤维连接蛋白结合。Western blotts分离到的病毒与高分子量质膜蛋白(S)的直接和特异性结合进一步证实了狂犬病病毒与这些细胞结合的蛋白受体的作用。
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-cotylglucoside 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 (∼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.