Molecular analysis of assembly and function of surface AMPA-receptor complexes in the mammalian brain

ampa型谷氨酸受体（AMPARs）是大脑兴奋性神经传递的核心参与者，是多蛋白复合物，其分子外观和功能特性由其蛋白质组（或相互作用组）编码，蛋白质组是其蛋白质构建块的集合。我们最初的蛋白质组学分析确定了30多种不同的蛋白质，这些蛋白质定义了内质网中“受体核心”的生物发生，以及“受体外围”的组装，最终在质膜的突触和突触外位点构建成熟的AMPAR复合物。事实上，我们最近的分析强烈表明，受体外周的成分是突触中ampar正常运作的主要决定因素，以及它们的活动依赖动力学和适应性，这是学习和记忆形成的基础。本研究旨在对表面AMPAR复合物（以“外周形成”相互作用组成分为特征）进行全面的分子理解，重点关注它们的组装、动力学、与突触蛋白的相互作用以及它们对受体介导的神经传递和可塑性的意义。为此，我们将采用跨学科的方法，包括生物化学和蛋白质组学分析，电生理学，以及荧光和电子显微镜。

Glutamate receptors of the AMPA-type (AMPARs), the central players of excitatory neurotransmission in the brain, are multi-protein complexes whose molecular appearance and functional properties are encoded by their proteome (or interactome), the ensemble of their protein building blocks. These more than 30 distinct proteins identified by our original proteomic analyses define the biogenesis of the ‘receptor-core’ in the endoplasmic reticulum, as well as the assembly of the ‘receptor periphery’ finally building the mature AMPAR complexes at synaptic and extra-synaptic sites of the plasma membrane. In fact, our recent analyses strongly suggest that the constituents of the receptor periphery are major determinants for proper operation of AMPARs in synapses, as well as their activity-dependent dynamics and adaptations that are fundamental for learning and memory formation. This proposal aims at a comprehensive molecular understanding of the surface AMPAR complexes (characterized by the ‘periphery forming’ interactome constituents), with focus on their assembly, their dynamics, their interactions with synaptic proteins and their significance for receptor-mediated neurotransmission and plasticity. For this purposes we will apply an interdisciplinary approach comprising biochemistry and proteomic analyses, electro-physiology, as well as fluorescence- and electron-microscopy.