小鼠不同组织的差异线粒体蛋白质组研究  

Proteomic Survey towards the Tissue-specific Proteins of Mouse Mitochondria

作　　者：王媛[1] 孙海丹[1] 茹雅维[1] 尹松月[1] 阴亮[1] 刘斯奇[1] 

机构地区：[1]中国科学院北京基因组研究所,北京101300

出　　处：《中国科学：生命科学》2010年第9期820-833,共14页Scientia Sinica(Vitae)

基　　金：国家高技术研究发展计划(批准号:2006AA02A308);国家自然科学基金(批准号:30700378)资助项目

摘　　要：线粒体是真核细胞的重要细胞器,它不仅具有合成ATP等普遍性功能,也具备一定的组织特异性,适应不同组织的生理功能需要.为了鉴定组织相关性线粒体蛋白,系统地分析和比较了C57BL/6J小鼠肝脏、肾脏和心脏线粒体蛋白质组.通过双向凝胶电泳和MALDI-TOF/TOF质谱,在这3种小鼠组织中共发现了87种组织特异的线粒体差异蛋白;采用ICPL的定量蛋白质组方法和Western blotting技术,进一步证实某些典型的组织特异性蛋白.还通过实时定量PCR和Western blotting技术分析了6种具有代表性的组织差异线粒体蛋白质在线粒体内外的分布,并结合激光共定位技术观察它们在相应组织活细胞内的表达与定位.研究发现,这些核基因编码的线粒体蛋白确有组织特异性分布和线粒体内外丰度差异特性.本结果为组织特异性线粒体蛋白质的深入研究奠定了实验基础.Mitochondrion plays the key functions in mammalian cells. It is believed that mitochondrion exerts the common biologic functions in many tissues, but also performs some specific functions correspondent with tissues where it is localized. To identify the tissue-specifically mitochondrial proteins, we carried out a systematic survey towards mitochondrial proteins in the tissues of C57BL/6J mouse, such as liver, kidney and heart. The mitochondrial proteins were separated by 2-DE and identified by MALDI-TOF/TOF MS. Total of 87 unique proteins were identified as the tissue-specific ones, and some representatives were further verified through ICPL quantification and Western blot. Since these tissue-specific proteins were all from the genes nucleic coded, we employed real-time PCR to examine the mRNA status of 6 typical genes in the tissues. With combining of the expression data and the co-localization images obtained from confocal microscope, we came to the conclusion that the tissue-specifically mitochondrial proteins were widely distributed along the mouse tissues. Our investigation, therefore, indeed provides a solid base to further explore the biological significance of the mitochondrial proteins with tissue-orientation.

关 键 词：蛋白质组学 组织特异性 线粒体 

分 类 号：Q593[生物学—生物化学]