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 遍布脊椎动物的神经系统。属性的更改 谷氨酸受体亚单位与以下疾病状态有关 癫痫、精神分裂症和短暂的兴奋性毒性细胞死亡 脑缺血，发生在中风中。谷氨酸的异构化 受体亚基决定了关键的受体特性，如钙 磁导率、电流幅度和电流-电压特性。 然而，亚基齐聚只发生在特定的谷氨酸中 受体亚家族，提示存在蛋白质结构域 赋予组装过程亚基专一性。长距离的 这些实验的目的是确定 测定谷氨酸受体亚基的寡聚化。具体目标#1 应用一种生化方法来定义和描述 参与亚基联合的GluR1亚基。在特定情况下的实验 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。这样的实验将会发现 亚基齐聚所必需的区域。为了确认这些信息 蛋白质结构域足以促进组装，嵌合通道将 在特定的目标3中构建，并测试他们的能力 与来自另一个子家族的亚单位相关联。最后，具体目标34 提出实验，以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.