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首页> 外文期刊>Science China Life Sciences >Antigenically dominant proteins within the human liver mitochondrial proteome identified by monoclonal antibodies
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Antigenically dominant proteins within the human liver mitochondrial proteome identified by monoclonal antibodies

机译：通过单克隆抗体鉴定的人肝线粒体蛋白质组中的抗原决定性蛋白

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Analysis of the mitochondrial proteome would provide valuable insight into the function of this important organelle, which plays key roles in energy metabolism, apoptosis, free radical production, thermogenesis, and calcium signaling. It could also increase our understanding about the mechanisms that promote mitochondrial disease. To identify proteins that are antigenically dominant in human liver mitochondria, we generated >240 hybridoma cell lines from native mitochondrial proteins after cell fusion, screening, and cloning. Antibodies that recognized mitochondrial proteins were identified by screening human liver cDNA expression libraries. In this study, we identified 6 major antigens that were recognized by at least 2 different monoclonal antibodies (mAbs). The proteins that were antigenically dominant were: acetyl-Coenzyme A acyltransferase 2 (mitochondrial 3-oxoacyl-Coenzyme A thiolase), aldehyde dehydrogenase 1 family member A1, carbamoyl phosphate synthetase 1, dihydrolipoamide S-acetyltransferase (E2 component of pyruvate dehydrogenase complex), enoyl coenzyme A hydratase 1, and hydroxysteroid (11-beta) dehydrogenase 1. We also determined the subcellular localizations of these enzymes within the mitochondria using immunohistocytochemistry. We believe that these well-characterized antibodies will provide a valuable resource for the Human Liver Proteome Project (HLPP), and will make studies aimed at investigating liver mitochondrial function far easier to perform in future. Our results provide strong evidence that, (i) depletion of dominant proteins from liver mitochondrial samples is possible and, (ii) the approaches adopted in this study can be used to explore or validate protein-protein interactions in this important organelle.
机译：线粒体蛋白质组的分析将提供对该重要细胞器功能的宝贵见解，该细胞器在能量代谢，细胞凋亡，自由基产生，生热和钙信号传导中起关键作用。它也可以增进我们对促进线粒体疾病机制的了解。为了鉴定在人肝线粒体中抗原性占主导地位的蛋白质，在细胞融合，筛选和克隆后，我们从天然线粒体蛋白质中生成了超过240种杂交瘤细胞系。通过筛选人类肝脏cDNA表达文库，鉴定出识别线粒体蛋白的抗体。在这项研究中，我们确定了至少2种不同的单克隆抗体（mAb）识别的6种主要抗原。抗原性占主导地位的蛋白质有：乙酰辅酶A酰基转移酶2（线粒体3-氧代酰基辅酶A硫解酶），醛脱氢酶1家族成员A1，氨基甲酰磷酸合成酶1，二氢脂酰胺S-乙酰转移酶（丙酮酸脱氢酶复合物的E2成分），烯酰辅酶A水合酶1和羟基类固醇（11-beta）脱氢酶1。我们还使用免疫组织化学测定了这些酶在线粒体内的亚细胞定位。我们相信，这些特性良好的抗体将为“人类肝蛋白质组计划”（HLPP）提供宝贵的资源，并将使旨在研究肝线粒体功能的研究将来更加容易进行。我们的结果提供了有力的证据，（i）可能从肝脏线粒体样品中去除了主要蛋白质，（ii）本研究中采用的方法可用于探索或验证该重要细胞器中的蛋白质-蛋白质相互作用。

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