Proteomic analysis of the presynaptic active zone from APP/APLP mutant mice

连接到突触前质膜的突触囊泡包括活跃区里具有胞吐能力的囊泡亚群。利用小鼠脑源性突触小体的亚细胞分离和针对整体囊泡膜蛋白的单克隆抗体，我们可以免疫分离突触囊泡及其附着的突触前质膜。APP家族的所有三个成员都存在于免疫分离的活性区部分中。我们的目标是比较APP/ aplp敲除小鼠与对照组突触前活性区的蛋白质组。此外，我们将研究在同窝对照和APP/APLP突变小鼠之间，诱导突触活动后突触前蛋白的翻译后修饰是否存在差异。蛋白质谱的差异将被分析，功能后果将由联盟内的共同努力评估。

Synaptic vesicles docked to the presynaptic plasma membrane comprise the exocytosiscompetent subpopulation of vesicles at the active zone. Employing subcellular fractionation of synaptosomes derived from mouse brain and a monoclonal antibody directed against an integral vesicle membrane protein, we could immunoisolate synaptic vesicles together with the attached presynaptic plasma membrane. All three members of the APP family are present in the immunoisolated active zone fraction. Our goal is to compare the proteome of the presynaptic active zone of APP/APLP-knockout mice to that of controls. In addition, we will investigate if there are differences in the posttranslational modifications of presynaptic proteins following induced synaptic activity between littermate controls and APP/APLP mutant mice. Differences in the protein profile will be analyzed and the functional consequences evaluated by a joint effort within the consortium.