EVIDENCE FOR MULTIPLE BINDING SITES FOR SEVERAL SPECIES OF HU IFN-ALPHA

几个 HU IFN-α 物种的多个结合位点的证据

• 批准号: 3804735
• 负责人: R Q HU
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3804735
• 关键词: clone cells; crosslink; cytokine receptors; interferons; lymphoblast; lymphocyte proliferation; molecular site; protein engineering; protein structure function; radiotracer; receptor binding; tissue /cell culture

The antiproliferative and competitive binding activities of several purified species of Hu lymphoblastoid Interferon-alpha were compared to that of recombinant Interferon-alpha 2b on Daudi and AU937 cells. We found that the antiproliferative activity of the interferon species on each cell line has a broad range and that Daudi cells were more sensitive than AU937 cells. The 50% inhibition of cell growth ranged from .003 ng/ml to greater than 10 ng/ml on Daudi cells and 1.2 ng/ml to >100 ng/ml on AU937 cells. Scatchard analyses demonstrated that the IFN-alpha 2b binding site number is greater on Daudi cells (12,700 binding sites/cell) than AU937 cells (3,300 binding sites/cell); however, their receptor affinities were similar. Interestingly, there appears to be three species (n,o,p) which exhibit the highest antiproliferative activities on both cell lines but do not compete for the Interferon-alpha-2b binding site. These data suggest that there may be more than one receptor or multiple receptor components involved in the biological actions of these molecules. In order to further study different receptors among species O IFN-alpha2b and other species, we have radiolabeled species O and performed competitive binding experiments. We also performed cross-linking to studies with IFN-alpha2b and species O to compare the molecular properties of the binding sites.

几种化合物的抗增殖和竞争性结合活性 将纯化的Hu类淋巴母细胞干扰素-α与 重组干扰素-α 2b对Daudi和AU 937细胞的作用。 我们 发现干扰素类的抗增殖活性对 每种细胞系都有广泛的范围，Daudi细胞更敏感， AU 937细胞 细胞生长的50%抑制范围为0.003 在Daudi细胞上为1.2ng/ml至>100 ng/ml， AU 937细胞 Scatchard分析表明，IFN-α 2b Daudi细胞上的结合位点数量更多（12，700个结合位点/细胞） 比AU 937细胞（3，300个结合位点/细胞）;然而，它们的受体 亲和力相似。 有趣的是，似乎有三种 （n，o，p），其在两种细胞上均表现出最高的抗增殖活性。 细胞系，但不竞争干扰素-α-2b结合位点。 这些数据表明，可能有一个以上的受体或多个 参与这些分子的生物学作用的受体成分。 为了进一步研究IFN-α 2b在物种间的不同受体， 和其他物种，我们放射性标记了物种O， 竞争性结合实验 我们还进行了交联， 使用IFN-α 2b和O类进行的比较分子特性的研究 的结合位点。