EXPRESSION AND MUTAGENESIS OF CLONED ALPHA2-RECEPTORS

目前的研究旨在检查，使用分子生物学 方法，结构基础的不同功能特性， α 2肾上腺素能受体 我们计划克隆猪的基因 我们已经纯化到同质的α 2肾上腺素能受体。 我们将利用从域映射和生物化学中获得的见解 分析纯化的受体以描绘受体的那些区域 我们可以通过缺失和定点突变来进一步分析 了解哪些结构成分参与配体识别， Na ~+、H ~+和5-氨基取代对肾上腺素能受体结合的变构效应 阿米洛利类似物，受体-GTP结合蛋白相互作用， 受体加速的Na+/H+交换和受体介导的抑制 腺苷酸环化酶 此外，我们计划研究什么角色不同， 受体的功能域在整个生理过程中起作用， 受α 2-肾上腺素能受体影响的功能，如抑制 神经递质释放，通过表达突变型与野生型受体 在适当的靶细胞中，以确定离散缺失或 位点突变具有特定的α 2受体功能。 我们预计 拟议的研究将提供新的见解， α 2受体的许多功能特性的基础， 提示药物开发的新位点，模拟或阻断， α 2-肾上腺素能药物在特定生理过程中的作用。

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.