MUSCARINIC RECEPTOR COUPLING MECHANISMS

毒蕈碱受体偶联机制

• 批准号: 3405936
• 负责人: GARY R LUTHIN
• 金额: $ 8.21万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3405936
• 关键词: adenylate cyclase; affinity chromatography; cholinergic agents; crosslink; gel electrophoresis; guanosine triphosphate; neuropharmacology; radionuclide double label

These studies will investigate the ability of muscarinic receptor proteins to interact with effector proteins. In membranes prepared from rat brain regions, interactions of muscarinic receptors with effector proteins will be studied using bifunctional protein-protein cross-linking agents, followed sequentially by solubilization, affinity chromatogrphy or immunoprecipitation, affinity labeling, and gel electrophoresis. The focus of these studies in membranes will be to establish conditions for the formation of covantly linked biospecific complexes of such species as receptor-Ni, which are known to modulate adenylate cyclase activity. Interaction of the muscarinic receptor with other effectors will be identified, in particular, recently identifiedd GTP-binding proteins. For these studies, membranes will be incubated with muscarinic agonists, and then with cross-linking agents. The covalent complexes thus formed will be solubilized and partially purified using affinity chromatograhy or immunoprecipitation. Fractions containing released proteins will be analyzed using gel electrophoresis. The identity of released proteins as IAP substrates and muscarinic ligand-binding proteins will be established following SDS-gel electrophoresis of covalently labelled proteins. These studies should provide useful information about muscarinic receptor-effector coupling mechanisms, as well as establish methodological protocols for examining muscarinic receptor-mediated responses.

这些研究将探讨毒蕈碱受体蛋白的能力