GLYCOPROTEIN HORMONE STRUCTURE

糖蛋白激素结构

• 批准号: 6240919
• 负责人: WILLIAM R MOYLE
• 金额: $ 21.77万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240919
• 关键词: Escherichia coli; SDS polyacrylamide gel electrophoresis; biological signal transduction; cell membrane; chickens; chimeric proteins; chorionic gonadotropin; dimer; follicle stimulating hormone; genetic mapping; glycoprotein structure; gonadotropins; hormone analog; hormone receptor; immunoprecipitation; luteinizing hormone; monoclonal antibody; oligosaccharides; protein purification; receptor binding; receptor expression; site directed mutagenesis; thyrotropin; tissue /cell culture

The overall objective of this project is to learn how the gonadotropins interact with their receptors to induce signal transduction. Previously we focused on identifying regions of the hormones that interact with their receptors. We identified portions of hCG and hFSH that control their abilities to distinguish LH and FSH receptors. Using an immunological strategy, we identified hormone residues that are exposed in the hormone receptor complex and could be detected by anti-hormone monoclonal antibodies. Combined with the crystal structure of deglycosylated hCG, these observations provide a model of how the hormone interacts with its receptor. Studies outlined in Specific Aims 1 and 2 will enable us to identify other portions of the hormone that are responsible for the receptor binding specificity of glycoprotein hormones. However, receptor binding is only the first event in signal transduction. While considerable information is available about the portions of the hormone that confer binding specificity, especially now that the crystal structure of deglycosylated hCG is known, much less is known about the events that occur to cause signal transduction. Aims 2 and 3 are designed to obtain information on the interactions of hCG with LH receptors that lead to signal transduction. Using a receptor analog that can be labeled, biotinylated analogs of hCG, and antibodies to the LH receptor, we will map the surfaces of hCG, deglycosylated hCG (an antagonist of hCG induced signal transduction), and the receptor. The studies employing biotinylated hCG should provide new information on hormone residues that are exposed after hCG binds to LH receptors. When applied to analogs of hCG that have hFSH activity, this technique should enable us to identify the different parts of the analog that interact with LH and FSH receptors. We are particularly interested in identifying specific portions of the hormone and the receptor that change following hormone binding and are associated with signal transduction. Therefore we will identify differences the portions of hCG and the receptor that are exposed after binding of hCG and deglycosylated hCG. Finally, we will obtain an independent estimate of the number of hormone and receptor molecules that are within the hormone receptor complex.

该项目的总体目标是了解促性腺激素如何 与其受体相互作用以诱导信号转导。之前 我们专注于识别与激素相互作用的区域 他们的受体。我们确定了控制 hCG 和 hFSH 的部分 他们区分 LH 和 FSH 受体的能力。使用 免疫学策略，我们鉴定了暴露的激素残留物 存在于激素受体复合物中，可以通过抗激素检测到 单克隆抗体。结合晶体结构 去糖基化 hCG，这些观察结果提供了激素如何 与其受体相互作用。具体目标 1 和 2 中概述的研究 将使我们能够识别激素的其他部分 负责糖蛋白的受体结合特异性 荷尔蒙。然而，受体结合只是信号中的第一个事件 转导。虽然有大量关于 赋予结合特异性的激素部分，尤其是现在 去糖基化 hCG 的晶体结构是已知的，更不用说了 了解引起信号转导的事件。目标2 和 3 旨在获取有关 hCG 与 导致信号转导的 LH 受体。使用受体类似物 可以标记的、生物素化的 hCG 类似物和抗体 LH 受体，我们将绘制 hCG、去糖基化 hCG（一种 hCG 诱导信号转导的拮抗剂）和受体。这 采用生物素化 hCG 的研究应提供以下新信息： hCG 与 LH 受体结合后暴露的激素残基。什么时候 应用于具有 hFSH 活性的 hCG 类似物，该技术应 使我们能够识别模拟中相互作用的不同部分 与 LH 和 FSH 受体。我们特别感兴趣的是识别 激素和受体的特定部分发生变化 激素结合并与信号转导相关。所以 我们将识别 hCG 和受体部分的差异 hCG 和去糖基化 hCG 结合后暴露。最后，我们将 获得激素和受体数量的独立估计 激素受体复合物内的分子。