Molecular mechanisms of glutamate transporters

描述（由申请人提供）：谷氨酸转运蛋白从突触中去除谷氨酸，以维持神经元之间的有效突触通信，并防止细胞外谷氨酸浓度达到神经毒性水平。本项目的长期目标是了解谷氨酸转运蛋白的分子机制。重要的是要了解谷氨酸转运蛋白实现谷氨酸摄取的分子机制，因为这将导致更好地理解大脑中的突触通讯，以及导致治疗，以防止由于谷氨酸毒性在例如缺血期间导致的神经元死亡。此外，这项研究将提供有关谷氨酸转运蛋白的基本结构和动力学信息，这是一个只有少数几个小组工作的研究领域。我们目前对谷氨酸转运体的结构和功能的了解非常有限。目前还没有结构模型可以解释谷氨酸转运蛋白是如何完成谷氨酸摄取的。我们建议使用荧光和生物化学方法来阐明谷氨酸转运蛋白的结构和功能，使用荧光和非荧光半胱氨酸反应探针。这些技术使我们能够获得谷氨酸转运蛋白的分子机制的结构和动力学信息。我们的目标是构建一个谷氨酸转运蛋白的结构模型，并阐明这种结构的变化如何导致谷氨酸转运。拟议项目的目的是：1）确定在转运周期中经历构象变化的谷氨酸转运蛋白的结构域; 2）确定在谷氨酸转运过程中经历交替访问的残基。

DESCRIPTION (provided by applicant): Glutamate transporters remove glutamate from synapses to maintain an efficient synaptic communication between neurons and to prevent extracellular glutamate concentrations from reaching neuro-toxic levels. The long-term goal of this project is to understand the molecular mechanism of glutamate transporter. It is important to understand the molecular mechanism by which glutamate transporters achieved glutamate uptake, because this will lead to a better understanding of synaptic communication in the brain, as well as, lead to treatments to prevent neuronal death due to glutamate toxicity during, for example, ischemia. In addition, this study will provide essential structural and dynamical information about the glutamate transporters, an area of research in which there are only a few numbers of groups working. Our current knowledge of the structure and function of glutamate transporters is very limited. No structural model has yet been suggested that can explain how glutamate transporters accomplish glutamate uptake. We propose to use fluorescence and biochemical methods to elucidate the structure and function of glutamate transporters, using fluorescent and non-fluorescent cysteine reactive probes. These techniques allow us to obtain structural as well as dynamical information about the molecular mechanism of glutamate transporters. Our objective is to construct a structural model of glutamate transporters and to elucidate how changes in this structure lead to glutamate transport. The aims of the proposed project are: 1) To identify the domains of the glutamate transporters that undergo conformational changes during the transport cycle; and 2) To identify residues that undergo alternating access during glutamate transport.