Yonally Sarah K, Capaldi Roderick A
Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229, United States.
Mitochondrion. 2006 Dec;6(6):305-14. doi: 10.1016/j.mito.2006.10.001. Epub 2006 Oct 14.
F(1)F(0) ATP synthase is ectopically expressed on the surface of several cell types, including endothelium and cancer cells. This study uses immunocytochemical detection methods via highly specific monoclonal antibodies to explore the possibility of plasma membrane localization of other mitochondrial proteins using an osteosarcoma cell line in which the location of the mitochondrial reticulum can be clearly traced by green fluorescent protein tagging of the organelle. We found that subunits of three of the four respiratory chain complexes were present on the surface of these cells. Additionally, we show for the first time that F(0) subunits d and OSCP of the ATP synthase are ectopically expressed. In all cases the OXPHOS proteins show a punctate distribution, consistent with data from proteome analysis of isolated lipid rafts that place the various mitochondrial proteins in plasma membrane microdomains. We also examined the cell surface for marker membrane proteins from several other intracellular organelles including ER, golgi and nuclear envelope. They were not found on the surface of the osteosarcoma cells. We conclude that mitochondrial membrane proteins are ectopically expressed, but not proteins from other cellular organelles. A specific mechanism by which the mitochondrion and plasma membrane fuse to deliver organellar proteins is suggested.
F(1)F(0) ATP合酶在包括内皮细胞和癌细胞在内的多种细胞类型表面异位表达。本研究使用免疫细胞化学检测方法,通过高度特异性单克隆抗体,利用骨肉瘤细胞系探索其他线粒体蛋白定位于质膜的可能性,在该细胞系中,通过对细胞器进行绿色荧光蛋白标记可清晰追踪线粒体网的位置。我们发现这四种呼吸链复合物中的三种亚基存在于这些细胞表面。此外,我们首次表明ATP合酶的F(0)亚基d和OSCP异位表达。在所有情况下,氧化磷酸化蛋白均呈点状分布,这与对分离的脂筏进行蛋白质组分析的数据一致,该分析将各种线粒体蛋白置于质膜微结构域中。我们还检测了骨肉瘤细胞表面是否存在来自其他几种细胞内细胞器(包括内质网、高尔基体和核膜)的标记膜蛋白。未在骨肉瘤细胞表面发现它们。我们得出结论,线粒体膜蛋白异位表达,但其他细胞器的蛋白并非如此。我们提出了一种线粒体与质膜融合以传递细胞器蛋白的特定机制。