Binding of 125I-labeled reovirus to cell surface receptors.
Binding of 125I-labeled reovirus to cell surface receptors.
复制标题
125I 标记的呼肠孤病毒与细胞表面受体的结合。
DOI：
10.1016/0042-6822(84)90424-0
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发表时间：
1984
期刊：
影响因子：
3.7
通讯作者：
Weiner,HL
中科院分区：
文献类型：
--
作者：
Epstein,RL;Powers,ML;Rogart,RB;Weiner,HL
定量研究了125 I标记的呼肠孤病毒与几种细胞类型的等量结合，包括（1）鼠L细胞成纤维细胞;（2）鼠脾T淋巴细胞;（3）YAC细胞，一种鼠淋巴瘤细胞系;和（4）R1.1细胞，一种鼠胸腺瘤细胞系。竞争性和饱和性研究表明：（1）呼肠孤病毒1型和3型与L细胞成纤维细胞上的不同受体特异性、可饱和、高亲和力结合;（2）呼肠孤病毒3型与小鼠脾淋巴细胞和R1.1细胞高亲和力结合;（3）呼肠孤病毒1型与淋巴细胞和R1.1细胞低亲和力结合;和（4）任一血清型与YAC细胞均无显著结合。发现两种呼肠孤病毒血清型与L细胞成纤维细胞的结合特征的差异是病毒血凝素的性质，如使用重组病毒克隆所证明的。病毒结合的平衡解离常数（Kd）具有极高的亲和力（Kin为0.5 nM），并且缓慢可逆。实验证明了呼肠孤病毒结合的温度和pH依赖性以及使用链霉蛋白酶、神经氨酸酶和各种糖的受体修饰研究证实了先前的研究，即呼肠孤病毒受体在结构上主要是蛋白质。呼肠孤病毒受体位点密度在2-8× 104个/细胞之间。这些研究表明，配体-受体相互作用的伪一级动力学模型为研究病毒颗粒与膜病毒受体的相互作用提供了一个有用的模型。他们还表明，一个细胞可能具有不同的受体位点，用于同一病毒的两种血清型，并且一种病毒血清型可能根据细胞类型以不同的动力学结合。
Quantitative studies of125I-labeled reovirus binding at equilbrium to several cell types was studied, including (1) murine L cell fibroblasts; (2) murine splenic T lymphocytes; (3) YAC cells, a murine lymphoma cell line; and (4) R1.1 cells, a murine thymoma cell line. Competition and saturation studies demonstrated (1) specific, saturable, high-affinity binding of reovirus types 1 and 3 to nonidentical receptors on L cell fibroblasts; (2) high-affinity binding of type 3 reovirus to murine splenic lymphocytes and R1.1 cells; (3) low-affinity binding of reovirus type 1 to lymphocytes and R1.1 cells; and (4) no significant binding of either serotype to YAC cells. Differences in the binding characteristics of the two reovirus serotypes to L cell fibroblasts were found to be a property of the viral hemagglutinin, as demonstrated using a recombinant viral clone. The equilibrium dissociation constant (Kd) for viral binding was of extremely high affinity (Kdin the range of 0.5 nM), and was slowly reversible. Experiments demonstrated temperature and pH dependence of reovirus binding and receptor modification studies using pronase, neuroaminidase, and various sugars confirmed previous studies that reovirus receptors are predominantly protein in structure. The reovirus receptor site density was in the range of 2–8×104sites/cell. These studies demonstrate that the pseudo-first-order kinetic model for ligand-receptor interactions provides a useful model for studying interactions of viral particles with membrane viral receptors. They also suggest that one cell may have distinct receptor sites for two serotypes of the same virus, and that one viral serotype may bind with different kinetics depending on the cell type.