Instituto de Bioquímica Vegetal y Fotosíntesis (IBVF), Centro de Investigaciones Científicas Isla de la Cartuja (cicCartuja), Universidad de Sevilla-Consejo Superior de Investigaciones Científicas (CSIC), Seville, 41092, Spain;
Mol Cell Proteomics. 2013 Dec;12(12):3666-76. doi: 10.1074/mcp.M113.030692. Epub 2013 Sep 9.
Programmed cell death is an event displayed by many different organisms along the evolutionary scale. In plants, programmed cell death is necessary for development and the hypersensitive response to stress or pathogenic infection. A common feature in programmed cell death across organisms is the translocation of cytochrome c from mitochondria to the cytosol. To better understand the role of cytochrome c in the onset of programmed cell death in plants, a proteomic approach was developed based on affinity chromatography and using Arabidopsis thaliana cytochrome c as bait. Using this approach, ten putative new cytochrome c partners were identified. Of these putative partners and as indicated by bimolecular fluorescence complementation, nine of them bind the heme protein in plant protoplasts and human cells as a heterologous system. The in vitro interaction between cytochrome c and such soluble cytochrome c-targets was further corroborated using surface plasmon resonance. Taken together, the results obtained in the study indicate that Arabidopsis thaliana cytochrome c interacts with several distinct proteins involved in protein folding, translational regulation, cell death, oxidative stress, DNA damage, energetic metabolism, and mRNA metabolism. Interestingly, some of these novel Arabidopsis thaliana cytochrome c-targets are closely related to those for Homo sapiens cytochrome c (Martínez-Fábregas et al., unpublished). These results indicate that the evolutionarily well-conserved cytosolic cytochrome c, appearing in organisms from plants to mammals, interacts with a wide range of targets on programmed cell death. The data have been deposited to the ProteomeXchange with identifier PXD000280.
程序性细胞死亡是沿进化尺度的许多不同生物体所表现出的一种事件。在植物中,程序性细胞死亡对于发育和对胁迫或致病感染的超敏反应是必要的。生物体中程序性细胞死亡的一个共同特征是细胞色素 c 从线粒体易位到细胞质。为了更好地理解细胞色素 c 在植物程序性细胞死亡中的作用,开发了一种基于亲和层析的蛋白质组学方法,并使用拟南芥细胞色素 c 作为诱饵。使用这种方法,鉴定出了十个假定的新细胞色素 c 伴侣。在这些假定的伴侣中,正如双分子荧光互补所表明的那样,其中 9 个在植物原生质体和人细胞中作为异源系统与血红素蛋白结合。使用表面等离子体共振进一步证实了细胞色素 c 与这种可溶性细胞色素 c 靶标之间的体外相互作用。总的来说,该研究获得的结果表明,拟南芥细胞色素 c 与涉及蛋白质折叠、翻译调节、细胞死亡、氧化应激、DNA 损伤、能量代谢和 mRNA 代谢的几个不同蛋白质相互作用。有趣的是,这些新的拟南芥细胞色素 c 靶标中的一些与人类细胞色素 c 的靶标密切相关(Martínez-Fábregas 等人,未发表)。这些结果表明,在从植物到哺乳动物的生物体中出现的进化上保守的细胞质细胞色素 c 与程序性细胞死亡过程中的广泛的靶标相互作用。这些数据已被存入 ProteomeXchange,标识符为 PXD000280。