Definition of the mitochondrial proteome by measurement of molecular masses of membrane proteins
Definition of the mitochondrial proteome by measurement of molecular masses of membrane proteins
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DOI：
10.1073/pnas.0607719103
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发表时间：
2006-10-31
影响因子：
11.1
通讯作者：
Walker, John E.
中科院分区：
文献类型：
--
作者：
Carroll, Joe;Fearnley, Ian M.;Walker, John E.
蛋白质的共价结构不完全由其基因序列定义，需要对完整蛋白质进行质谱分析以检测任何翻译后修饰的存在。由于大多数膜蛋白是在与质谱电离技术不相容的去污剂中纯化的，因此许多疏水蛋白没有进行这种必要的测量，因此蛋白质组学数据不完整。我们已经提取了膜蛋白从牛线粒体和洗涤剂纯化的NADH：泛醌氧化还原酶（复合物1）与有机溶剂，分馏的混合物通过亲水相互作用色谱法，并测量了完整的膜蛋白的分子量，包括6个亚基的复合物I的编码在线粒体DNA。这些测量解决了线粒体基因组解释的长期不确定性，并且它们对复合物I的共价组成的定义做出了重大贡献。
The covalent structure of a protein is incompletely defined by its gene sequence, and mass spectrometric analysis of the intact protein is needed to detect the presence of any posttranslational modifications. Because most membrane proteins are purified in detergents that are incompatible with mass spectrometric ionization techniques, this essential measurement has not been made on many hydrophobic proteins, and so proteomic data are incomplete. We have extracted membrane proteins from bovine mitochondria and detergent-purified NADH:ubiquinone oxidoreductase (complex 1) with organic solvents, fractionated the mixtures by hydrophilic interaction chromatography, and measured the molecular masses of the intact membrane proteins, including those of six subunits of complex I that are encoded in mitochondrial DNA. These measurements resolve long-standing uncertainties about the interpretation of the mitochondrial genoe, and they contribute significantly to the definition of the covalent composition of complex I.