THYROTROPIN RELEASING HORMONE RECEPTOR MOLECULAR BIOLOGY

促甲状腺素释放激素受体分子生物学

• 批准号: 2824954
• 负责人: MARVIN C GERSHENGORN
• 金额: $ 1.44万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2824954
• 关键词: G protein; computer simulation; conformation; fluorescence microscopy; hormone receptor; hormone regulation /control mechanism; hydrogen bond; immunofluorescence technique; immunoprecipitation; inositol phosphates; intermolecular interaction; mutant; phosphorylation; physical model; pituitary gland; protein sequence; protein structure function; receptor binding; receptor coupling; receptor expression; receptor mediated endocytosis; site directed mutagenesis; structural biology; thyrotropin releasing hormone; tissue /cell culture

The broad, long term objective of this research program is to define at a molecular level the mechanism(s) and regulation of signal transduction by thyrotropin-releasing hormone (TRH) and its GTP-binding regulatory (G) protein-coupled receptor (TRH-R). These studies will lead to a better understanding of cell regulation under physiologic and pathologic circumstances by TRH and all hormones, growth factors and neurotransmitters that signal via receptors of this family. The findings may lead to rational design of new drugs to treat diseases of these signalling systems. This proposal is timely because new insights into these diseases are now being gained and a combined approach using molecular biology and computer modelling holds great promise for a deeper understanding of these processes. Three interrelated aspects of TRH-R biology will be explored. The Specific Aims are: #1. To model the 3-D structure of TRH-R. 1A. To further define the atomic interactions between TRH and TRH-R. 1B. To further define the intramolecular interactions among transmembrane helices of TRH-R. 1C. To develop a model of the extracellular loops of TRH-R and determine whether these loops serve to direct TRH into the binding pocket. #2. To delineate the mechanism of TRH- R activation. 2A. To identify the G protein coupling domain(s) of TRH-R and determine the specific amino acids involved in receptor/G protein coupling. 2B. To identify transmembrane residues in TRH-R that are important for activation. 2C. To develop a computer simulation of the conformational changes in TRH-R that are associated with transition from the inactive to the activated state. #3. To identify the sequences in TRH- R that mediate the post-translational processes of internalization and downregulation, and to delineate these mechanisms. For Specific Aims #1 and #2, an interactive approach using tools of molecular biology to experimentally analyze structure-function of TRH-R will be combined with computer simulations that are based on the experimental observations and will guide further experimentation. For Specific Aim #3, mutagenesis of TRH-R will be combined with cell biologic studies. Experiments will be performed by gene transfer of native and mutated proteins into cell culture systems.

该研究计划的广泛长期目标是在 分子水平的机制和信号转导的调节 促甲状腺蛋白释放激素（TRH）及其GTP结合调节（G） 蛋白质偶联受体（TRH-R）。这些研究将导致更好 在生理和病理学下了解细胞调节 TRH和所有激素，增长因素和 通过该家族的受体发出信号的神经递质。发现 可能会导致新药的理性设计来治疗这些疾病 信号系统。该建议是及时的，因为新见解 这些疾病现在正在获得，并使用 分子生物学和计算机建模具有巨大的希望 了解这些过程。 TRH-R的三个相互关联的方面 将探索生物学。具体目的是：＃1。建模3-D TRH-R的结构。 1a。进一步定义了原子相互作用 TRH和TRH-R。 1B。进一步定义分子内相互作用 TRH-R的跨膜螺旋。 1C。开发一个模型 TRH-R的细胞外环，并确定这些环是否用于 将TRH直接进入装订袋。 ＃2。描绘trh-的机制 R激活。 2a。确定TRH-R的G蛋白偶联域 并确定受体/G蛋白涉及的特异性氨基酸 耦合。 2b。识别TRH-R中的跨膜残基 对激活很重要。 2C。开发计算机模拟 与从 对激活状态的无效。 ＃3。识别trh-中的序列 R调解了内部化后翻译后的过程 下调，并描述这些机制。对于特定目标＃1 ＃2，一种使用分子生物学工具的交互式方法 实验分析TRH-R的结构功能将与 基于实验观察的计算机模拟和 将指导进一步的实验。对于特定目的＃3， TRH-R将与细胞生物学研究结合使用。实验将是 通过将天然和突变蛋白基因转移到细胞中进行的 培养系统。