The Potato Tuber Mitochondrial Proteome
The Potato Tuber Mitochondrial Proteome
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DOI：
10.1104/pp.113.229054
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发表时间：
2014-02-01
期刊：
影响因子：
7.4
通讯作者：
Moller, Ian Max
中科院分区：
文献类型：
--
作者：
Salvato, Fernanda;Havelund, Jesper F.;Moller, Ian Max
线粒体被称为细胞的发电站。为了更好地了解线粒体在维持和调节贮藏组织代谢中的作用，从休眠马铃薯块茎（Solanum tuberosum 'Folva'）中分离纯化了高纯度的线粒体，并对其蛋白质组进行了研究。通过一维凝胶电泳解析蛋白质，并从凝胶切片中提取胰蛋白酶肽，并使用Orbitrap XL通过液相色谱-串联质谱法进行分析。使用四种不同的搜索程序，共1,060个非冗余的蛋白质被确定在定量的方式使用归一化的光谱计数，包括多达5倍的“极端”蛋白质（低质量，高等电点，疏水性）比以前的线粒体蛋白质组研究。我们估计，这个蛋白质的纲要代表了马铃薯块茎线粒体蛋白质组的高覆盖率（可能高达85%）。蛋白质表达的动态范围跨越1,800倍，几乎包括电子传递链、三羧酸循环和蛋白质输入装置的所有组分。此外，我们确定了71 pentatricopeptide重复蛋白，29膜载体/转运蛋白，一些新的蛋白参与辅酶生物合成和铁代谢，丙酮酸脱氢酶激酶，和2C型蛋白磷酸酶，可能催化丙酮酸脱氢酶复合物的去磷酸化。系统分析的显着的翻译后修饰显示，超过50%的已确定的蛋白质港口至少有一个修改。最显著的翻译后修饰是氧化修饰。这项研究揭示了大约500种新的或以前未经证实的植物线粒体蛋白，并概述了一种简单的策略，用于无偏见的，近乎全面的线粒体蛋白及其修饰形式的鉴定。
Mitochondria are called the powerhouses of the cell. To better understand the role of mitochondria in maintaining and regulating metabolism in storage tissues, highly purified mitochondria were isolated from dormant potato tubers (Solanum tuberosum 'Folva') and their proteome investigated. Proteins were resolved by one-dimensional gel electrophoresis, and tryptic peptides were extracted from gel slices and analyzed by liquid chromatography-tandem mass spectrometry using an Orbitrap XL. Using four different search programs, a total of 1,060 nonredundant proteins were identified in a quantitative manner using normalized spectral counts including as many as 5-fold more "extreme" proteins (low mass, high isoelectric point, hydrophobic) than previous mitochondrial proteome studies. We estimate that this compendium of proteins represents a high coverage of the potato tuber mitochondrial proteome (possibly as high as 85%). The dynamic range of protein expression spanned 1,800-fold and included nearly all components of the electron transport chain, tricarboxylic acid cycle, and protein import apparatus. Additionally, we identified 71 pentatricopeptide repeat proteins, 29 membrane carriers/transporters, a number of new proteins involved in coenzyme biosynthesis and iron metabolism, the pyruvate dehydrogenase kinase, and a type 2C protein phosphatase that may catalyze the dephosphorylation of the pyruvate dehydrogenase complex. Systematic analysis of prominent posttranslational modifications revealed that more than 50% of the identified proteins harbor at least one modification. The most prominently observed class of posttranslational modifications was oxidative modifications. This study reveals approximately 500 new or previously unconfirmed plant mitochondrial proteins and outlines a facile strategy for unbiased, near-comprehensive identification of mitochondrial proteins and their modified forms.