MEMBRANE EVENTS FOLLOWING HORMONE BINDING TO LH RECEPTOR

激素与 LH 受体结合后的膜事件

• 批准号: 2888948
• 负责人: Deborah A Roess
• 金额: $ 12.95万
• 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 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2888948
• 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.

黄体激素（LH）受体在叶黄素和Leydig上 睾丸中的细胞结合黄体生成激素（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受体与其他膜成分的相互作用以及 可产生信号的激素受体复合物的细胞骨架 转导。首先，我们将检查LH受体的相互作用 与其他能够激活的细胞中的其他质膜蛋白 响应激素结合的腺苷酸环化酶。我们将比较 结合激素但无法的LH受体的分子动力学 与功能性LH受体复合物的传递信号。 其次，我们将检查与该膜相互作用的特定膜蛋白 配体结合后的LH受体，尤其是那些可能是 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以检查受体 受体相互作用和光活化的膜探针以识别 受体附近的膜蛋白。第三，我们将确定如何 细胞骨架限制LH的运动，通过限制受体 在亚微米内膜结构域中。我们将检查横向扩散 以及在存在的野生型和突变体LH受体的轨迹 缺乏使用光漂白恢复方法完整的细胞骨架 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 和细胞膜上存在的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 细胞中磷脂酶C的受体未用 密集表达的LH受体能够激活营地和 磷脂酶C.科罗拉多州立大学的融合 生殖生理学和工具设施的专业知识 膜结构研究。这种情况为我们提供了独特的 研究LH受体组织以及如何研究的机会 组织调节受体功能。