Biophysical characterization of chemokine receptors

描述(由申请人提供)： 这项提议主要目的是回答几个尚未解决的基本问题 关于谷氨酸转运体的结构和功能的问题。一个 已经提出了一系列实验来表征低聚物 功能转运蛋白的结构并阐明其结构 与亚基组装和底物选择性有关的特征。具体目标1 将表征谷氨酸转运体的寡聚体结构 利用传输器子类型之间的差异来映射所需的域 用于亚单位组装。此外，神经元特异性异源多聚体 运输商将接受检查。这是一个关键的实验，因为它一直是 研究表明，EAAT3和EAAT4神经元转运蛋白在脑内共同表达 小脑浦肯野细胞。诱变与生化技术 本提案中描述的氨基酸残基将确定 转运体之间的底物选择性。在特定目标3中，细菌 谷氨酸转运体将被提纯以获得X射线衍射物 水晶。谷氨酸转运蛋白分子机制的研究进展 功能是理解突触传递的关键。

DESCRIPTION (as provided by Applicant): The main goal of this proposal is to answer several unresolved fundamental questions regarding the structure and function of glutamate transporters. A series of experiments have been proposed to characterize the oligomeric structure of the functional transporters and to elucidate the structural features involved in subunit assembly and substrate selectivity. Specific aim 1 will characterize the oligomeric structure of glutamate transporters and will utilize the differences between transporter subtypes to map domains necessary for subunit assembly. In addition, heteromultimerization of neuronal specific transporters will be examined. This is a critical experiment since it has been shown that EAAT3 and EAAT4 neuronal transporters are expressed together in the Purkinje cells of cerebellum. The mutagenesis and biochemical techniques described in this proposal will determine the amino acid residues necessary for substrate selectivity among transporters. In specific aim 3, bacterial glutamate transporter will be purified in order to obtain x-ray diffractable crystals. Elucidating the molecular mechanisms underlying glutamate transporter function is essential in understanding synaptic transmission.