Molecular mechanisms of glutamate transporters

谷氨酸转运蛋白的分子机制

• 批准号: 7020638
• 负责人: Hans Peter Larsson
• 金额: $ 30.03万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7020638
• 关键词: Xenopus oocyte; binding sites; confocal scanning microscopy; conformation; cysteine; fluorescence microscopy; fluorescence resonance energy transfer; fluorimetry; glutamate transporter; model design /development; neural transmission; neurotransmitter transport; physical model; protein protein interaction; site directed mutagenesis; synapses

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.

描述（由申请人提供）：谷氨酸转运蛋白从突触中去除谷氨酸，以保持神经元之间有效的突触通信，并防止细胞外谷氨酸浓度达到神经毒性水平。该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，因为这将使人们更好地理解大脑突触通信，并导致治疗以防止因缺血期间因谷氨酸毒性而导致的神经元死亡。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，该研究领域只有少数几组工作。我们目前对谷氨酸转运蛋白的结构和功能的了解非常有限。尚未提出任何可以解释谷氨酸转运蛋白如何完成谷氨酸摄取的结构模型。我们建议使用荧光和非荧光半胱氨酸的反应性探针使用荧光和生化方法来阐明谷氨酸转运蛋白的结构和功能。这些技术使我们能够获得有关谷氨酸转运蛋白分子机制的结构和动力信息。我们的目标是构建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）确定在运输周期中发生构象变化的谷氨酸转运蛋白的域； 2）确定在谷氨酸运输过程中可以交替进入的残留物。