肝细胞线粒体-脂滴互作的分子机制研究

越来越多的研究表明，线粒体-脂滴互作参与调控脂质代谢过程，对于维持肝脏组织的脂质代谢稳态至关重要，但我们对于参与线粒体-脂滴互作的蛋白及其作用机制仍然知之甚少。本项目将结合双分子荧光互补技术和基于APEX2蛋白的邻近标记策略，建立新的、系统的线粒体-脂滴互作位点蛋白质组的鉴定分析方法，并利用这一方法比较正常条件和葡萄糖饥饿条件下肝细胞线粒体-脂滴互作位点蛋白质组的差异，绘制动态蛋白质组图谱。本项目的实施有助于发现新的、关键的定位于线粒体-脂滴互作位点的蛋白质，揭示其分子机制，为调控脂质代谢提供新的靶标。此外，本项目建立的方法具有很强的可拓展性，可以被应用到任意的细胞器互作位点，这将有利于细胞器互作领域的研究。

More and more studies have shown that mitochondria-lipid droplet interaction is involved in the regulation of lipid metabolism and crucial for maintaining the homeostasis of lipid metabolism in liver. However, the proteins involved in mitochondria-lipid droplet interaction and the functioning mechanism remain largely elusive. In this project, we will develop a novel method to systematically identify the proteome of mitochondria-lipid droplet contact site by combining the bimolecular fluorescence complementary assay and proximity labeling strategy based on APEX2. Then we plan to use this method to compare the proteome of mitochondria-lipid droplet contact site in hepatic cells under normal condition and glucose deprivation condition. The implementation of this project will help to discover new and key proteins located at mitochondria-lipid droplet contact site, reveal its molecular mechanism and provide new therapeutic targets for the regulation of lipid metabolism. In addition, the strategy used in this project could be applied to any other inter-organelle contact site, which will benefit the research of inter-organelle interaction.