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MUSCARINIC RECEPTOR COUPLING MECHANISMS

毒蕈碱受体偶联机制

基本信息

批准号：
3405938
负责人：
GARY R LUTHIN
金额：
$ 8.96万
依托单位：
ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-07-01 至 1992-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3405938
关键词：
adenylate cyclase biological signal transduction brain mapping enzyme mechanism guanine nucleotide binding protein inhibitor /antagonist laboratory rabbit laboratory rat muscarinic receptor neuropharmacology receptor coupling receptor expression stimulant /agonist

项目摘要

The long-term objectives of this project are to study mechanisms of signal transduction in the brain. A first step to understanding this process is t develop biochemical methodologies allowing molecular characterization of receptor and effector entities. To this end, antibodies to neurotransmitte receptors and guanine nucleotide binding proteins (G proteins) have been developed, taking advantage of recent elucidation of their primary amino ac d sequences. The antibodies,are used in an immunoaffinity purification of biologically active complexes of muscarinic receptor and G protein. Specifically, muscarinic receptors are labelled in cellular membranes with agonists or antagonists, then solubilized from the membranes. The receptor are purified over a lectin affinity resin, then immunoprecipitated with anti-receptor antibodies. In the presence of agonist, this allows co-purification of associated G proteins. The G proteins present in comple with the receptor are identified using monospecific antibodies to their subunits. This experimental design will be used to identify muscarinic receptor-G protein complexes in various brain regions, and is of interest because of the recent cloning of at least 5 subtypes of the receptor, most of which can be found in any of several brain regions. The experiments can resolve the issue of which G protein each receptor subtype interacts with i a given brain region. The combination of different receptor subtypes, and a potential of each subtype to interact with any of several G proteins, cou d be the molecular basis for the apparent specificity of cellular responses t receptor agonists. A dysfunction of cellular signal transduction could occ r as a result of inappropriate expression of a receptor or G protein subtype, with a consequent subordination off normal signal pathways. This could in turn form the basis for disease states such as Altzheimer's, and for learni g and memory deficits where the cholinergic pathways in brain are believed to be altered.

这个项目的长期目标是研究信号的机制。大脑中的转导。理解这一过程的第一步是开发生物化学方法，允许分子表征受体和效应器实体。为此，针对神经递质的抗体受体和鸟嘌呤核苷酸结合蛋白(G蛋白)已被利用最近对它们的主要氨基酸酯的阐明而开发的 D 序列。这些抗体用于免疫亲和纯化 M受体与G蛋白的生物活性复合体。具体地说，M受体在细胞膜上用激动剂或拮抗剂，然后从膜上溶解。感受器通过凝集素亲和树脂纯化，然后免疫沉淀抗受体抗体。在激动剂存在的情况下，这允许联合纯化相关G蛋白。G蛋白以复杂的形式存在使用针对其受体的单特异性抗体进行鉴定亚单位。这一实验设计将被用来鉴定毒鼠强受体-G蛋白复合体在不同的大脑区域，并引起人们的兴趣由于最近克隆了至少5种受体亚型，大多数它们可以在几个大脑区域中的任何一个中找到。这些实验可以解决每个受体亚型与哪种G蛋白相互作用的问题一个给定的大脑区域。不同受体亚型的组合，以及每种亚型与几种G蛋白相互作用的潜力，例如 D 是细胞反应表观特异性的分子基础受体激动剂。细胞信号转导功能障碍可OCC R 由于受体或G蛋白亚型表达不当，从而使其脱离正常的信号通路。这可能会在形成阿尔茨海默氏症等疾病状态的基础，以及学习障碍的基础 G 以及记忆缺陷，大脑中的胆碱能通路被认为是被改变。