MOLECULAR BIOLOGY OF NEUROTRANSMITTER RECEPTORS

神经递质受体的分子生物学

• 批准号: 3922594
• 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/3922594
• 关键词: Chondrichthyes; Drosophilidae; X ray crystallography; benzodiazepines; beta adrenergic receptor; biological polymorphism; complementary DNA; developmental genetics; gamma aminobutyrate; gene expression; genetic transduction; glycine; human tissue; laboratory rat; molecular biology; muscarinic receptor; neurophysiology; nicotinate; protein structure; serotonin; site directed mutagenesis; species difference

This project is to characterize the gene and protein structure and evolution of neurotransmitter receptors belonging to two major multi gene families. The gene families are those containing adrenergic, muscarinic, opsin, seratonin receptors and the second family containing nicotinic cholinergic, GABA/benzodiazepine and glycine receptors. The specific aims are to clone and sequence the genes for receptors in these two multigene families; to obtain very high density receptor expression and to use the expressed proteins to determine the complete structure of these receptors by x-ray crystallography and computer enhanced molecular modeling; to determine the evolution of the neurotransmitter receptor gene families; to search for and characterize receptor gene polymorphisms. To date we have cloned and sequenced a significant number of receptor genes from multigene families. These include beta 2-adrenergic receptors from human brain and genomic library, rat cardiac and rat genomic library and from shark genomic library. Beta 1-adrenergic receptors from a human genomic library; M1-M4 muscarinic cholinergic receptors from human, rat, shark and Drosophila genomic library. The human alpha-2 adrenergic receptor from a human genomic library; human alpha and beta subunits of the GABA/benzodiazepine receptor from human genomic libraries; adrenergic receptors from Drosophila cDNA and genomic library; nicotinic receptor genes from neuronal cDNA library and from locust cDNA libraries, and a chick genomic clone of the GABA benzodiazepine receptor. These receptors have been cloned into expression vectors and mammalian cells transfected. The results are permanent cell lines expressing the unique neurotransmitter receptor protein. The expressed receptor lines are providing key new information concerning the mechanism of receptors activated by neurotransmitters and a basis of receptor structural determinants.

该项目旨在描述基因和蛋白质结构， 神经递质受体的进化属于两个主要的 多基因家族 基因家族是那些包含 肾上腺素能受体、毒蕈碱受体、视蛋白受体、血清素受体和第二受体 含烟碱胆碱能、GABA/苯二氮卓类和 甘氨酸受体 具体目标是克隆和测序 这两个多基因家族中的受体基因;为了获得非常 高密度受体表达和使用表达的蛋白质 通过X射线确定这些受体的完整结构 晶体学和计算机增强分子建模; 确定神经递质受体基因的进化 寻找和鉴定受体基因 多态性 到目前为止，我们已经克隆和测序了一个重要的 来自多基因家族的受体基因的数量。 这些包括 来自人脑和基因组文库的β 2-肾上腺素能受体， 大鼠心脏和大鼠基因组文库以及鲨鱼基因组文库。 人类基因组文库中的β 1-肾上腺素能受体; M1-M4 来自人、大鼠、鲨鱼和 果蝇基因组文库。 人α-2肾上腺素能受体 来自人类基因组文库; 人基因组文库中GABA/苯二氮卓受体的研究 果蝇肾上腺素能受体cDNA和基因组文库; 神经元cDNA文库和蝗虫烟碱受体基因 cDNA文库和GABA的鸡基因组克隆 苯二氮卓受体 这些受体已经被克隆到 表达载体和转染的哺乳动物细胞。 结果 是表达独特神经递质的永久细胞系 受体蛋白 表达的受体细胞系提供了 关于受体激活机制的新信息 神经递质和受体结构决定因素的基础。