MOLECULAR BASIS OF NON-NMDA GLUTAMATE RECEPTOR ASSEMBLY

离子型谷氨酸受体介导快速兴奋性神经传递 在脊椎动物的神经系统中。的属性变化 谷氨酸受体亚单位与这些疾病状态有关， 癫痫性精神分裂症，以及短暂性 缺血，这发生在中风中。谷氨酸的异源多聚化 受体亚单位决定了受体的关键特性， 磁导率、电流幅度和电流-电压特性。 然而，亚基寡聚化仅发生在某些谷氨酸内， 受体亚家族，表明存在蛋白质结构域 赋予组装过程亚基特异性。远程 这些实验的目的是确定分子机制， 确定谷氨酸受体亚单位的寡聚化。具体目标#1 应用生物化学方法来定义和表征 GluR 1亚基参与亚基缔合。具体实验 目的#2评估这些蛋白质结构域在组装过程中的作用 使用电生理分析。这些实验将确定 亚基寡聚化所必需的区域。为了证实这些 蛋白质结构域足以促进组装，嵌合通道将 在具体目标#3中构建并测试其能力， 与来自另一个亚家族的亚基结合。第34章具体目标 提出实验来探索动力学和相对亲和力， 使用生物分子相互作用分析的亚基相互作用。

Ionotropic glutamate receptors mediate fast excitatory neurotransmission throughout the vertebrate nervous system. Changes in the properties of glutamate receptor subunits has been implicated in such disease states as epilepsy schizophrenia, and in excitotoxic cell death following transient ischemia, which occurs in stroke. Heteromultimerization of glutamate receptor subunits determines critical receptor properties such as calcium permeability, current amplitude, and current-voltage characterization. Subunit oligomerization, however, occurs only within certain glutamate receptor subfamilies, suggesting the existence of a protein domain conferring subunit specificity on the assembly process. The long-range objective of these experiments is to define the molecular mechanisms that determine oligomerization of glutamate receptor subunits. Specific Aim #1 applies a biochemical approach to define and characterize regions of the GluR1 subunit involved in subunit associations. Experiments in Specific Aim #2 evaluate the roles of such protein domains in the assembly process using electrophysiological analyses. Such experiments will identify regions necessary for subunit oligomerization. In order to confirm these protein domains are sufficient to promote assembly, chimeric channels will be constructed in Specific Aim #3 and tested for their ability to associate with subunits from another subfamily. Finally, Specific Aim 34 proposes experiments to explore the kinetics and relative affinities of subunit interactions using biomolecular interaction analyses.