MEMBRANE EVENTS FOLLOWING HORMONE BINDING TO LH RECEPTOR

激素与 LH 受体结合后的膜事件

• 批准号: 6182044
• 负责人: Deborah A Roess
• 金额: $ 14.33万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6182044
• 关键词: G protein; SDS polyacrylamide gel electrophoresis; adenylate cyclase; biological signal transduction; chorionic gonadotropin; cytoskeleton; fertility; hormone receptor; hormone regulation /control mechanism; immunoprecipitation; luteinizing hormone; membrane activity; membrane proteins; phospholipase C; protein structure function; radioimmunoassay; receptor binding; receptor expression; sheep; single cell analysis; statistics /biometry; tissue /cell culture; western blottings

Luteinizing hormone (LH) receptors on the corpus luteum and on Leydig cells in the testis bind luteinizing hormone (LH) and human chorionic gonadotropin (hCG) to regulate steroid synthesis and secretion from these glands. Our basic hypothesis is that hormone binding is accompanied by interactions of the LH receptor with other membrane components and with the cytoskeleton to produce hormone-receptor complexes capable of signal transduction. First, we will examine the interactions of the LH receptor with other plasma membrane proteins in cells capable of activating adenylate cyclase in response to hormone binding. We will compare the molecular dynamics of the LH receptors that bind hormone but are unable to transduce signal with those of functional LH receptor complexes. Second, we will examine specific membrane proteins that interact with the LH receptor following ligand binding, particularly those which might be present only when formation of the hormone-receptor complex leads to activation of adenylate cyclase. We will use biophysical methods including fluorescence energy transfer techniques to examine receptor- receptor interactions and photoactivated membrane probes to identify membrane proteins near the receptor. Third, we will determine how the cytoskeleton restricts the movement of LH, possible by confining receptor within sub-micron membrane domains. We will examine the lateral diffusion and trajectories of wild-type and mutant LH receptors in the presence and absence of an intact cytoskeleton using photobleaching recovery methods and single particle tracking techniques. Fourth, we will determine whether activation of phospholipase C is dependent on the organization and interactions of LH receptors present on the cell membrane. We will compare the molecular motions and interactions of sparsely expressed LH receptors in cells where phospholipase C is not activated with those of densely expressed LH receptors capable of activating both cAMP and phospholipase C. Colorado State University has an unusual confluence of expertise in reproductive physiology and instrumental facilities for membrane structural studies. This situation provides us a unique opportunity to study both LH receptor organization and how this organization modulates receptor function.

黄体和间质上的黄体生成素 (LH) 受体 睾丸细胞结合黄体生成素 (LH) 和人绒毛膜 促性腺激素 (hCG) 调节类固醇的合成和分泌 腺体。我们的基本假设是激素结合伴随着 LH 受体与其他膜成分以及与 细胞骨架产生能够发出信号的激素-受体复合物 转导。首先，我们将检查 LH 受体的相互作用 与细胞中其他质膜蛋白能够激活 腺苷酸环化酶响应激素结合。我们将比较 结合激素但不能结合的 LH 受体的分子动力学 用功能性 LH 受体复合物转导信号。 其次，我们将检查与 配体结合后的 LH 受体，特别是那些可能 仅当激素-受体复合物的形成导致 腺苷酸环化酶的激活。我们将使用生物物理方法 包括荧光能量转移技术来检查受体 受体相互作用和光敏膜探针来识别 受体附近的膜蛋白。第三，我们将确定如何 细胞骨架通过限制受体来限制 LH 的运动 在亚微米膜域内。我们将检查横向扩散 以及存在和存在时野生型和突变型 LH 受体的轨迹 使用光漂白恢复方法缺乏完整的细胞骨架 和单粒子跟踪技术。第四，我们将确定 磷脂酶C的激活是否取决于组织 以及细胞膜上 LH 受体的相互作用。我们将 比较稀疏表达的 LH 的分子运动和相互作用 磷脂酶 C 不被细胞中的受体激活 密集表达的 LH 受体能够激活 cAMP 和 磷脂酶 C. 科罗拉多州立大学有一个不寻常的融合 生殖生理学方面的专业知识和仪器设施 膜结构研究。这种情况为我们提供了独特的 有机会研究 LH 受体组织及其如何 组织调节受体功能。