生物钟基因与代谢之间具有明显相关性。我们前期研究发现部分生物钟CLOCK蛋白分布于线粒体，并与线粒体膜转运蛋白SLC25A10之间存在相互作用。故提出假说，CLOCK蛋白可能通过乙酰化SLC25A10，影响了线粒体膜内外物质转运来调节线粒体代谢功能，进而影响整个机体的代谢水平。我们首先将利用蛋白质pull-down技术、Surface Plasma Resonance技术进一步验证二者的结合能力，找到SLC25A10上的结合位点和乙酰化位点；利用代谢组学方法研究点突变SLC25A10细胞内转运相关底物的改变，利用海马XF系统检测细胞线粒体呼吸代谢功能改变；最后，在体外建立SLC25A10结合序列突变和乙酰化位点突变的小鼠，研究其对线粒体代谢的重要作用和相关表型，并在分子水平上进一步验证上述作用的机制。本课题将为研究生物钟基因调节线粒体与物质和能量代谢提供新的实验基础。

There is a close correlation between the circadian genes and metabolism. Our previous study found that circadian CLOCK protein located in the mitochondria and interacted with a mitochondrial carrier protein SLC25A10. Therefore, we propose that CLOCK can bind to SLC25A10 and acetylate SLC25A10, thereby regulate mitochondrial respiratory function and eventually affect the body's energy metabolism. We will first use protein pull-down technology, surface plasma resonance technology to verify the binding capacity of the two proteins as well as find the key acetylation sites in SLC25A10 through the point mutation method. Metabolomics will be subsequently used to study the changes of SLC25A10 transportation related substrates. The Seahorse facility will be used to study the mitochondrial respiratory metabolism and functional changes of various mitochondrial respiratory complexes. Finally, we will establish SLC2510A point mutant mice, metabolic cages and other methods will be used to study the impact of mutations on mitochondrial function and metabolism in vivo to clarify the above mechanism. Our study will provide an important experimental basis for the regulation roles of circadian gene clock in mitochondrial metabolism.