Structure and Function of the LH, FSH, and TSH Receptors

LH、FSH 和 TSH 受体的结构和功能

• 批准号: 7068040
• 负责人: DEBORAH SEGALOFF
• 金额: $ 41.24万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7068040
• 关键词: G protein; SDS polyacrylamide gel electrophoresis; biological signal transduction; follicle stimulating hormone; gene mutation; hormone receptor; luteinizing hormone; molecular site; protein structure function; receptor binding; receptor coupling; thyrotropin; western blottings

DESCRIPTION (provided by applicant): The LH, FSH and TSH receptors, collectively known as the glycoprotein hormone receptor (GPH-R's), comprise a unique subfamily of rhodopsin-like G protein-coupled receptors (GPCR's). They are each composed of a large extracellular domain that binds hormone and a seven transmembrane (TM) domain that couples to G proteins, primarily Gs. The active states of each of these receptors are stabilized by the binding of hormone or by discrete mutations that induce constitutive activity. The aims of this grant are to determine the structural basis for the activation of the GPH-R's. In general, we will use two experimental approaches, each of which will be coupled with molecular modeling. The first approach utilizes disruptive and reciprocal mutagenesis to test the hypothesis that TM residues that are highly conserved in the three GPH-R's participate in interhelical interactions that stabilize the receptors in the resting state. The second approach uses selective chimera mutagenesis between different GPH-R's from a given species or from a given GPH-R of different species to test the hypothesis that certain divergent TM residues modulate the activities of the GPH-R's by affecting interhelical interactions. In spite of the high degree of amino acid identity in the TM regions of the GPH-R's, certain pairs of GPH-R's exhibit markedly different degrees of basal activity, mutation-induced, or hormone-induced activation. By interchanging the divergent TM residues between two related GPH-R's with differing properties, we can maintain the overall integrity of the receptor (because the divergent residues are capable of maintaining the overall structure) and determine the precise amino acids that confer the greater or lesser degree of activity. Computer modeling will be coupled with each of these experimental strategies to aid in data interpretation and to define those interhelical interactions that stabilize the GPH-R's in the resting states, in mutation-induced active states, and in hormone-stabilized active states. The specific aims of this proposal are: 1. Determine the roles that conserved residues of the GPH-R's have in maintaining the resting states of the receptors. 2. Determine the structural basis for the different degrees of constitutive activity of the resting states of the GPH-R's. 3. Determine the structural basis for the different susceptibilities of the GPH-R's to be stabilized in an active state by activating mutations. 4. Determine the structural basis for the different degrees of hormone-stimulated Gs activation by GPH-R's.

描述（由申请人提供）：LH、FSH和TSH受体，统称为糖蛋白激素受体（GPH-R），包括一个独特的视紫红质样G蛋白偶联受体（GPCR）亚家族。它们各自由结合激素的大细胞外结构域和与G蛋白（主要是Gs）偶联的七个跨膜（TM）结构域组成。这些受体中的每一种的活性状态通过激素的结合或通过诱导组成型活性的离散突变而稳定。该补助金的目的是确定激活GPH-R的结构基础。一般来说，我们将使用两种实验方法，每种方法都将与分子建模相结合。第一种方法利用破坏性和相互诱变来测试以下假设：在三个GPH-R中高度保守的TM残基参与使受体稳定在静息状态的螺旋间相互作用。第二种方法使用来自给定物种的不同GPH-R之间或来自不同物种的给定GPH-R之间的选择性嵌合诱变来检验某些不同TM残基通过影响螺旋间相互作用来调节GPH-R活性的假设。尽管GPH-R ′ s的TM区的氨基酸高度相同，但某些GPH-R ′ s对表现出明显不同程度的基础活性、突变诱导的或突变诱导的活化。通过在具有不同性质的两个相关GPH-R之间互换不同的TM残基，我们可以保持受体的整体完整性（因为不同的残基能够保持整体结构）并确定赋予更大或更小程度活性的精确氨基酸。计算机建模将与这些实验策略中的每一个相结合，以帮助数据解释，并定义那些稳定GPH-R在静息状态，突变诱导的活性状态，并在稳定的活性状态的螺旋间的相互作用。该提案的具体目标是：1.确定GPH-R的保守残基在维持受体静息状态中的作用。2.确定GPH-R的静息状态的组成活性的不同程度的结构基础。3.确定GPH-R的不同亲和性的结构基础，以通过激活突变稳定在活性状态。4.确定GPH-R对葡萄糖刺激的Gs激活的不同程度的结构基础。