Structural Characterization of Muscarinic and Alpha Adrenergic Receptors

毒蕈碱和α肾上腺素受体的结构表征

• 批准号: 9817140
• 负责人: George Turner
• 金额: $ 19万
• 依托单位: University of Miami School of Medicine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9817140&HistoricalAwards=false
• 关键词: Structural; Characterization; Muscarinic; Alpha; Adrenergic

The objective of these studies is to understand the structure and function of two important membrane signaling receptors: the muscarinic acetylcholine (subtype M1) receptor and the alpha 2B adrenergic receptor. Both are members of a large family of structurally related G protein-coupled receptors. The experimental approach is to use a novel protein expression system with the potential of generating each receptor in the quantities large enough to permit many standard structural and functional analyses that have not yet been possible. The approach entails the construction of chimeras of the genes encoding the receptors and the bacteriorhodopsin gene to direct high level expression in the halophilic archaeon Halobacterium salinarum. Screening of chimeras with different portions of the bacteriorhodopsin gene will define an optimal strategy for receptor over-expression. In addition, constructs will also be expressed in previously employed eukaryotic cell systems as a means to test receptor function in vivo. The method will ensure that the successfully expressed chimeras retain their ability to couple to endogenous downstream signaling components. The studies have a high potential to yield significant new information about the molecular mechanisms of signaling by these membrane proteins.

这些研究的目的是了解两个重要的膜信号受体的结构和功能：毒蕈碱乙酰胆碱（亚型M1）受体和α2B肾上腺素能受体。 两者都是大型结构相关的G蛋白偶联受体的成员。 实验方法是使用一种新型的蛋白质表达系统，其潜力是在足够大的数量中产生每个受体，以允许许多尚未实现的标准结构和功能分析。 该方法需要建立编码受体和细菌紫红素基因的基因的嵌合体，以指导盐谷盐盐盐盐中的高水平表达。 用不同部分的细菌蛋白基因筛选嵌合体将定义受体过表达的最佳策略。 此外，在先前使用的真核细胞系统中，构建体也将作为在体内测试受体功能的一种手段。 该方法将确保成功表达的嵌合体保持其夫妇为内源性下游信号传导组件的能力。 这些研究具有很高的潜力，可以产生有关这些膜蛋白信号传导的分子机制的重要新信息。