CHARACTERIZATION OF HEART ATRIAL MUSCARINIC RECEPTOR

心房毒蕈碱受体的表征

• 批准号: 3337339
• 负责人: Michael Schimerlik
• 金额: $ 22.56万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-12-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3337339
• 关键词: G protein; atrium; biological signal transduction; circular dichroism; conformation; fluorescent dye /probe; heart pharmacology; ligands; muscarinic receptor; protein purification; protein reconstitution; protein structure function; receptor binding; receptor coupling; site directed mutagenesis; stop flow technique; swine; tissue /cell culture; ultraviolet spectrometry

Muscarinic cholinergic receptors in the atria mediate the regulation of the heart by the parasympathetic nervous systems. In order to obtain detailed information regarding the molecular mechanism of atrial muscarinic receptor signal transduction two approaches are proposed. The first is to use site- directed mutagenesis to explore the roles of specific amino acids in ligand binding, thermal stability and coupling to either endogenous Chinese Hamster ovary (CHO) cell G proteins or to atrial Gi reconstituted into CHO cell membranes. The second approach is to use biophysical methods such as circular dichroism and transient kinetics to examine changes in secondary structure that occur when ligands bind to the purified recombinant protein or to detect and analyze protein conformational changes that occur in the membrane-bound or purified preparations. Interaction of defined Gialpha1,2,3, with atrial Gibetagamma and muscarinic receptors, by resolving purified atrial gi into gil and Gi3 or by purifying Gi1,2,3 from brain and erythrocytes. The detailed knowledge of muscarinic mechanisms in the heart may be useful in the rational design of drugs for use in treating cardiac arrhymias as tachycardia.

心房中的毒蕈碱胆碱能受体介导了心房肌细胞的 心脏的副交感神经系统。为了获得详细的 心房毒蕈碱受体的分子机制 提出了两种信号转导途径。第一个是使用网站- 定向突变以探索配体中特定氨基酸的作用 结合、热稳定性和偶联到内源性中国 卵巢（CHO）细胞G蛋白或心房Gi重构为CHO 细胞膜第二种方法是使用生物物理方法，例如 圆二色性和瞬态动力学，以检查次级 当配体与纯化的重组蛋白结合时出现的结构 或者检测和分析在细胞中发生的蛋白质构象变化， 膜结合的或纯化的制剂。定义的相互作用 Gialpha 1，2，3，心房Gibetagamma和毒蕈碱受体， 将纯化的心房gi分解为gil和Gi 3，或通过从 脑和红细胞。详细了解毒蕈碱机制， 心脏可用于合理设计用于治疗 心律失常如心动过速。