粟酒裂殖酵母中Mmp1蛋白的功能研究  

Studies on the Function of Mmp1 Protein in Schizosaccharomyces Pombe

作　　者：赵兰 黄鹰[1] Zhao Lan;Huang Ying(School of Life Sciences,Nanjing Normal University,Nanjing 210023,China)

机构地区：[1]南京师范大学生命科学学院,江苏南京210023

出　　处：《南京师范大学学报（工程技术版）》2022年第4期69-74,共6页Journal of Nanjing Normal University(Engineering and Technology Edition)

基　　金：国家自然科学基金项目(31400012).

摘　　要：线粒体是存在于大多数真核细胞中的一种产能细胞器,它参与了细胞内ATP的合成、物质的代谢、细胞凋亡等多种生理过程.Mmp1蛋白被预测为线粒体内膜肽酶复合体催化亚基,但其具体功能尚未得到解析.通过分子克隆、表型实验、荧光显微观察、线粒体分离提取、实时荧光定量PCR、蛋白质印迹法等技术,分析了Mmp1的定位及其对线粒体功能的影响.通过荧光定位分析发现Mmp1是一个定位在线粒体中的蛋白.细胞缺失mmp1无法在非发酵型培养基上生长,暗示缺失mmp1影响了线粒体的功能.同时,细胞缺失mmp1,使线粒体基因组编码的呼吸链复合体核心蛋白Cox1、Cox2、Cox3、Cob1、Atp6的蛋白水平显著降低.在对野生型和Δmmp1菌株线粒体编码的基因的mRNA水平进行实时荧光定量实验发现,Δmmp1菌株中的Cox1、Cox2、Cox3、Cob1、Atp6的转录水平并未有明显变化.结果表明,Mmp1影响了线粒体呼吸链复合体亚基的蛋白水平,对维持线粒体功能的发挥起着重要作用.Mitochondria are the productive organelles of most eukaryotic cells,which are involved in various physiological processes such as ATP synthesis,substance metabolism and cell apoptosis.Mmp1 protein is predicted to be the catalytic subunit of the mitochondrial endopeptidase complex,but its specific function has not been analyzed.In this paper,the localization of Mmp1 and its effect on mitochondrial function are analyzed by molecular cloning,phenotypic experiment,fluorescence microscopic observation,mitochondrial isolation and extraction,real-time fluorescence quantitative PCR and western-blot.Fluorescence localization analysis reveal that Mmp1 is a protein located in mitochondria.Δmmp1 strain can not grow on the medium with glycerol as carbon source and can not perform aerobic respiration,suggesting that the deletion of mmp1 affects mitochondrial function.Meanwhile,the protein levels of Cox1,Cox2,Cox3,Cob1 and Atp6,the core proteins of respiratory chain complex encoded by mitochondrial genome,are significantly reduced when mmp1 is knock out.The transcriptional levels of Cox1,Cox2,Cox3,Cob1 and Atp6 inΔmmp1 strain do not change significantly.These results indicate that Mmp1 affects the protein levels of some mitochondrial respiratory chain complex subunits and plays an important role in maintaining mitochondrial function.

关 键 词：MMP1 线粒体 实时荧光定量PCR 线粒体蛋白提取 

分 类 号：Q78[生物学—分子生物学]