The Arabidopsis thaliana 2-D gel mitochondrial proteome: Refining the value of reference maps for assessing protein abundance, contaminants and post-translational modifications
The Arabidopsis thaliana 2-D gel mitochondrial proteome: Refining the value of reference maps for assessing protein abundance, contaminants and post-translational modifications
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DOI：
10.1002/pmic.201000620
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发表时间：
2011-05-01
期刊：
影响因子：
3.4
通讯作者：
Millar, A. Harvey
中科院分区：
文献类型：
--
作者：
Taylor, Nicolas L.;Heazlewood, Joshua L.;Millar, A. Harvey
线粒体进行氧化磷酸化过程，产生植物细胞维持和生长所需的三磷酸腺苷。几十年来，植物线粒体蛋白质的分离和分级原理已经得到了改进，并对许多植物物种的线粒体蛋白质组进行了调查。随着时间的推移，通过二维凝胶分析对植物线粒体蛋白质组的变化进行了许多定量分析，揭示了线粒体蛋白质在对突变、胁迫和发育的反应中的诱导、降解和修饰。在这里，我们提出了一种饱和的MS分析，从一个典型的拟南芥线粒体纯化的二维凝胶可分离蛋白质点识别264个蛋白质，同时用非凝胶方法的LC-MS/MS调查识别220个蛋白质。这使得我们能够表征在二维凝胶上没有观察到的主要线粒体蛋白质、常见污染物和关键线粒体生化途径的蛋白质机制的丰富性，并认为N末端前序列裂解和磷酸化的影响可以解释多个蛋白质点和蛋白质在二维凝胶上的配位。
Mitochondria undertake the process of oxidative phosphorylation yielding ATP for plant cell maintenance and growth. The principles of isolation and fractionation of plant mitochondrial proteins have been improved over decades, and surveys of the mitochondrial proteome in a number of plants species have been performed. Over time, many quantitative analyses of changes in the plant mitochondrial proteome have been performed by 2-D gel analyses revealing the induction, degradation and modification of mitochondrial proteins in responses to mutation, stress and development. Here, we present a saturating MS analysis of 2-D gel separable protein spots from a typical purification of Arabidopsis mitochondria identifying 264 proteins, alongside an LC-MS/MS survey by non-gel methods identifying 220 proteins. This allowed us to characterise the major mitochondrial proteins that are not observed on 2-D gels, the common contaminants and the abundance of the protein machinery of key mitochondrial biochemical pathways, and consider the impact of N-terminal pre-sequence cleavage and phosphorylation as explanations of multiple protein spots and the co-ordinates of proteins on 2-D gels.