MAJOR HISTOCOMPATIBILITY ANTIGENS & MEMBRANE RECEPTOR

我们的目标是显示MHC抗原对配体的影响程度 与多肽激素受体结合，进而被修饰为 配体与特定受体结合的结果。我们已经显示为 人类白细胞抗原(基因和表型)与亲和力的关系 B淋巴母细胞的胰岛素受体。我们现在将进一步分析 人类白细胞抗原与胰岛素受体亲和力和延展性的遗传关联 抗原性与抗原性的物理关系分析 受体分子。基因分析将通过检测进行 突变的淋巴母细胞，因表达某些人类白细胞抗原而缺失，这 已经被导入了缺失抗原的基因。我们还将 尝试挽救人类白细胞抗原在Daudi细胞中的表达，以检测 这种抢救对Daudi与胰岛素结合的影响。物理分析 将采取两种方法：1.尝试，基于 文献，共沉淀胰岛素受体，通过他们的检测 内源性酪氨酸激酶活性，与I类MHC抗原，使用 特异性抗受体和抗I类单抗及2。 I类MHC抗原与胰岛素接近度的测定 完整细胞中的受体，利用共振能量转移 内源性(Beta-2-m)标记的I类抗原和胰岛素受体 我们已经展示了它可以用生物素胰岛素和 亲和素藻胆蛋白。能量转移将在流动中被测量 流式细胞仪在供体猝灭受体敏化发射方面。 我们计划扩展我们在表皮生长的代谢影响方面的工作。 因子结合对野生型和野生型的展示和磷酸化状态的影响 突变的第I类MHC抗原，使用流式细胞术来定量显示 以及标准的免疫化学和生化方法来确定 与细胞周期变化相关的抗原的磷酸化状态 展示。

Our goal is to show the extent to which MHC antigens influence ligand binding by peptide hormone receptors and in turn are modified as a consequence of ligand binding to specific receptors. We have shown as association between HLA (genotype and phenotype) and the affinity of insulin receptors on B lymphoblasts. We will now further analyze the genetic associations between HLA and insulin receptor affinity and extend the analysis to the physical relationships between the antigens and receptor molecules. The genetic analysis will proceed through examination of mutant lymphoblasts, deleted for expression of some HLA antigens, which have been transfected with genes for the missing antigens. We will also attempt to rescue expression of HLA antigens in Daudi cells, to examine the effects of such rescue on insulin binding by Daudi. The physical analysis will take two approaches: 1. an attempt, based on precedents in the literature, to co-precipitate insulin receptors, detected by their endogenous tyrosine kinase activity, with class I MHC antigens, using specific anti-receptor and anti-class I monoclonal antibodies and 2. determination of the proximity of class I MHC antigens and insulin receptors in intact cells, using resonance energy transfer between endogenously (Beta-2-m) labeled class I antigens, and insulin receptors which we have shown can be specifically labeled with biotinyl insulin and avidin phycobilliproteins. Energy transfer will be measured in the flow cytometer in terms of donor quenching sensitized emission of acceptor. We plan to extend our work on the metabolic effects of epidermal growth factor binding on the display and phosphorylation state of wild-type and mutant class I MHC antigens, using flow cytometry to quantitate the display and standard immunochemical and biochemical approaches to determine the phosphorylation state of the antigens associated with changes in the display.