MOLECULAR BIOLOGY OF NEUROTRANSMITTER RECEPTORS

神经递质受体的分子生物学

• 批准号: 3881766
• 负责人: J C VENTER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3881766
• 关键词: Caenorhabditis elegans; Chondrichthyes; Drosophilidae; GABA receptor; NMDA receptors; Orthoptera; X ray crystallography; adrenergic receptor; benzodiazepine receptor; beta adrenergic receptor; biochemical evolution; biological polymorphism; chickens; clone cells; complementary DNA; computer simulation; gene expression; genetic library; genetic mapping; genetic polymorphism; genome; glycine receptors; human tissue; laboratory rat; molecular biology; molecular cloning; molecular genetics; muscarinic receptor; neurotransmitter receptor; nicotinic receptors; nucleic acid sequence; nucleic acid structure; octopamine; physical model; protein sequence; protein structure; receptor expression; serotonin receptor; species difference; transfection

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.

该项目旨在表征基因和蛋白质结构以及 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、 血清素和章鱼胺受体，第二个包括烟碱受体 胆碱能受体、GABA/苯二氮卓类 NMDA 和甘氨酸受体。具体的 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并使用 表达的蛋白质部分确定完整的受体结构 通过计算机增强分子建模；来确定进化的 神经递质受体基因家族；并寻找和表征 受体基因多态性。迄今为止，我们已经克隆并测序了 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 的多种 α 和 β 肾上腺素能受体 和基因组文库；来自人类、大鼠和小鼠的毒蕈碱胆碱能受体 果蝇基因组文库。果蝇的章鱼胺受体；人类 GABA/苯二氮卓受体的α和β亚基；和烟碱的 来自蝗虫和线虫的受体。例如人类 GABA beta 1 亚基已被克隆、测序并定位于 4 号染色体。 基因超过65kb，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，并且较低 物种。表达独特神经递质的永久细胞系 受体蛋白正在提供有关 神经递质激活受体的机制。