MOLECULAR BIOLOGY OF NEUROTRANSMITTER RECEPTORS

该项目旨在描述基因和蛋白质结构， 属于两个主要多基因的神经递质受体的进化 家庭第一个基因家族包括肾上腺素能、毒蕈碱、视蛋白， 第一种包括5-羟色胺和章鱼胺受体，第二种包括烟碱受体， 胆碱能、GABA/苯并二氮杂NMDA和甘氨酸受体。具体 目的是克隆和测序这两种受体的基因， 多基因家族;获得高密度受体表达和应用 表达的蛋白质，以确定部分完整的受体结构， 通过计算机增强的分子建模;以确定 神经递质受体基因家族;并寻找和表征 受体基因多态性到目前为止，我们已经克隆和测序了一个 来自两个多基因家族的大量受体基因。这些 包括来自人和大鼠cDNA几种α和β-肾上腺素能受体 和基因组文库;来自人、大鼠和 果蝇基因组文库。果蝇章鱼胺受体;人 GABA/苯二氮卓受体的α和β亚基;以及烟碱 蝗虫和C.优雅例如，人类GABA β 1 亚基已被克隆和测序并定位于4号染色体。这 基因大小超过65 kb，由9个外显子组成。外显子和剪接位点 在人类的其他GABA受体亚基中高度保守， 物种表达独特神经递质的永久细胞系 受体蛋白质提供了关键的新信息 神经递质激活受体的机制。

This project is to characterize the gene and protein structure and evolution of neurotransmitter receptors belonging to two major multigene families. The first gene family comprises adrenergic, muscarinic, opsin, serotonin and octopamine receptors and the second comprises nicotinic cholinergic, GABA/benzodiazepine NMDA, and glycine receptors. The specific aims are to clone and sequence the genes for receptors in these two multigene families; to obtain high density receptor expression and to use the expressed proteins to determine the complete receptor structure in part by computer enhanced molecular modeling; to determine the evolution of the neurotransmitter receptor gene families; and to search for and characterize receptor gene polymorphisms. To date, we have cloned and sequenced a significant number of receptor genes from both multigene families. These include several alpha and beta-adrenergic receptors from human and rat cDNA and genomic libraries; muscarinic cholinergic receptor from human, rat and Drosophila genomic libraries. Octopamine receptors for Drosophila; human alpha and beta subunits of the GABA/benzodiazepine receptor; and nicotinic receptors from locusts and C. elegans. For example the human GABA beta 1 subunit has been cloned and sequenced and localized to chromosome 4. This gene is over 65kb and is composed of nine exons. The exons and splice sites are highly conserved in other GABA receptor subunits from humans and lower species. Permanent cell lines expressing the unique neurotransmitter receptor proteins are providing key new information concerning the mechanism of receptors activation by neurotransmitters.