PINK1蛋白与BAG2蛋白的相互作用在帕金森病发病机制中的作用研究

BAG2蛋白是我们在前期工作中运用酵母双杂交系统筛选出来的PINK1蛋白的相互作用蛋白，并应用免疫共沉淀及体外结合实验证实BAG2蛋白与PINK1蛋白存在相互作用，且存在直接相互作用。本项目在此研究基础上，利用体外细胞模型和PINK1突变的果蝇模型，运用免疫荧光共定位、RNA干扰、免疫共沉淀、体外蛋白结合实验、放射自显影等.分子生物学和蛋白质功能研究技术，进一步证实PINK1蛋白与BAG2蛋白的相互作用，探讨PINK1基因R492X突变对PINK1蛋白与BAG2蛋白的相互作用及亚细胞分布的影响，确定BAG2蛋白对野生型和突变型PINK1蛋白的稳定性、泛素系统降解及线粒体功能的影响，明确野生型和突变型PINK1蛋白能否磷酸化修饰BAG2蛋白，探讨HSP70对BAG2-PINK1调节作用的影响，从分子水平深入探讨PINK1蛋白与BAG5蛋白的相互作用在帕金森病发病机制中的作用。

BAG2 is one of PINK1-interaction-proteins screened by yeast two-hybrid system in our previous study. We have identified that BAG2 can interact directly with PINK1. Based on these results, we will use cell cultures and a PINK1 mutation drosophila model , and a series of molecular biological techniques and protein functional study techniques such as immunofluorescence co-localization, RNAi, co-immunoprecipitation, protein binding test in vitro and radioautography. Our project will aim to explore the influence of PINK1 R492X mutation on interaction between them and subcellular location, determine the role of BAG2 in the stability of PINK1, degradation of ubiquitin system and mitochondrial function, identify whether PINK1 can phosphorylate and modify BAG2 and investigate the influence of HSP70 on BAG2-PINK1 interaction, explore the role of interaction between PINK1 and BAG2 in the pathogenesis of Parkinson's disease at molecular level.