MEMBRANE EVENTS FOLLOWING HORMONE BINDING TO LH RECEPTOR

黄体和Leydig上的促黄体激素（LH）受体 睾丸中的细胞结合促黄体生成激素（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受体与其他膜组分以及与 细胞骨架产生能够传递信号的受体复合物 转导首先，我们将研究LH受体的相互作用， 与细胞中的其他质膜蛋白能够激活 腺苷酸环化酶对激素结合的反应。我们将比较 LH受体的分子动力学结合激素，但不能 与功能性LH受体复合物的信号传导相一致。 第二，我们将研究特定的膜蛋白， LH受体与配体结合后，特别是那些可能 只有当受体-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学方法 包括荧光能量转移技术来检测受体- 受体相互作用和光活化膜探针， 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，可能是通过限制受体 在亚微米膜区域内。我们将研究横向扩散 和野生型和突变型LH受体的轨迹， 使用光漂白恢复方法的完整细胞骨架缺失 和单粒子跟踪技术。第四，我们将确定 磷脂酶C的激活是否依赖于组织 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不被那些 密集表达的LH受体能够激活cAMP和 脂酶C已科罗拉多州立大学有一个不寻常的融合， 生殖生理学和仪器设备方面的专门知识， 膜结构研究这种情况为我们提供了一个独特的 有机会研究LH受体的组织和如何， 组织调节受体功能。

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.