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首页> 外文期刊>Proteomics >Quantitative and qualitative 2D electrophoretic analysis of differentially expressed mitochondrial proteins from five mouse organs
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Quantitative and qualitative 2D electrophoretic analysis of differentially expressed mitochondrial proteins from five mouse organs

机译：五个小鼠器官中差异表达的线粒体蛋白质的定量和定性二维电泳分析

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Mitochondria fulfill many tissue-specific functions in cell metabolism. We set out to identify differences in the protein composition of mitochondria from five tissues frequently affected by mitochondrial disorders. The proteome of highly purified mitochondria from five mouse organs was separated by high-resolution 2DE. Tissue-specific spots were identified through nano-LC/ESI-MS/MS and quantified by densitometry in ten biological replicates. We identified 87 consistently deviating spots representing 48 proteins. The percentage of variant spots ranged between 4.2% and 6.0%; 21 proteins having tissue-specific isospots. Consistent tissue-specific processing/regulation was seen for carbamoyl-phosphate-synthase, aldehyde-dehydrogenase 2, ATP-synthase α-chain, and isocitrate-dehydrogenase α-subunit. Thirty tissue-specific proteins were associated with mitochondrial disorders in humans. We further identified alcohol-dehydrogenase, catalase, quinone-oxidoreductase, cyclophilin-A, and Upf0317, a potential biotin-carboxyl-carrier protein, which had not been annotated as "mitochondrial" in Gene Ontology or MitoCarta databases. Their targeting to the mitochondria was verified by transfection of full-length GFP-tagged plasmids. Given the high evolutionary conservation of mitochondrial metabolic pathways, these data further annotate the mitochondrial proteome and advance our understanding of the pathophysiology and tissue-specificity of symptoms seen in patients with mitochondrial disorders. The generation of 2D electrophoretic maps of the mitochondrial proteome using tissue specimens in the milligram range facilitates this technique for clinical applications and biomarker research.
机译：线粒体在细胞代谢中具有许多组织特有的功能。我们着手从经常受线粒体疾病影响的五个组织中鉴定线粒体蛋白质组成的差异。来自五个小鼠器官的高纯度线粒体蛋白质组被高分辨率2DE分离。通过nano-LC / ESI-MS / MS鉴定组织特异性斑点，并通过光密度法对十个生物学重复进行定量。我们确定了代表48种蛋白质的87个一致偏离的斑点。变异点的百分比在4.2％至6.0％之间;具有组织特异性同位点的21种蛋白质。氨基甲酸酯磷酸合酶，醛脱氢酶2，ATP合酶α链和异柠檬酸脱氢酶α亚基的组织特异性加工/调节一致。三十种组织特异性蛋白质与人类线粒体疾病有关。我们进一步确定了醇脱氢酶，过氧化氢酶，醌氧化还原酶，亲环蛋白-A和Upf0317（一种潜在的生物素-羧基载体蛋白），在基因本体论或MitoCarta数据库中未标注为“线粒体”。通过靶向全长GFP标签的质粒，证实了它们靶向线粒体。鉴于线粒体代谢途径的高度进化保守性，这些数据进一步注释了线粒体蛋白质组，并增进了我们对线粒体疾病患者所见症状的病理生理和组织特异性的了解。使用毫克范围内的组织标本生成线粒体蛋白质组的二维电泳图谱有助于将该技术用于临床应用和生物标记物研究。

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