EXPRESSION AND MUTAGENESIS OF CLONED ALPHA2-RECEPTORS

克隆的 α2 受体的表达和诱变

• 批准号: 2637969
• 负责人: LEE E LIMBIRD
• 金额: $ 25.84万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2637969
• 关键词: adenylate cyclase; adrenergic receptor; computer assisted sequence analysis; enzyme inhibitors; eukaryote; gene deletion mutation; guanosine triphosphate; hydrogen channel; immunoprecipitation; ion exchange chromatography; ion transport; molecular cloning; molecular genetics; nucleic acid sequence; protein structure function; receptor binding; receptor expression; site directed mutagenesis; sodium channel; swine; transfection

The present studies are aimed at examining, using a molecular biological approach, the structural basis for the diverse functional properties of alpha2-adrenergic receptors. We plan to clone the porcine gene that codes for the alpha2-adrenergic receptor that we have purified to homogeneity. We will exploit insights gained from domain mapping and biochemical analysis of the purified receptor to delineate those areas of the receptor that we can analyze further by deletion and site-directed mutagenesis to learn what structural components are involved in ligand recognition, allosteric effects on adrenergic binding by Na+, H+ and 5-amino-substituted analogs of amiloride, receptor-GTP binding protein interactions, receptor-accelerated Na+/H+ exchange and receptor mediated inhibition of adenylate cyclase. Furthermore, we plan to examine what role different functional domains of the receptor play in the overall physiological functions influenced by alpha2-adrenergic receptors, such as suppression of neurotransmitter release, by expressing mutated versus wild type receptors in appropriate target cells to determine what effect discrete deletion or site mutations have on particular alpha2-receptor functions. We anticipate that the proposed studies will provide new insights into the structural basis for many of the functional properties of alpha2-receptors and perhaps suggest novel loci for drug development in mimicking, or blocking, the effects of alpha2-adrenergic agents in particular physiological processes.

目前的研究目的是利用分子生物学的方法进行检测