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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Comparative proteomics of human endothelial cell caveolae and rafts using two-dimensional gel electrophoresis and mass spectrometry
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Comparative proteomics of human endothelial cell caveolae and rafts using two-dimensional gel electrophoresis and mass spectrometry

机译：使用二维凝胶电泳和质谱法比较人内皮细胞小窝和筏的蛋白质组学

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摘要

The human endothelial cell plasma membrane harbors two subdomains of similar lipid composition, caveolae and rafts, both crucially involved in various essential cellular processes like transcytosis, signal transduction and cholesterol homeostasis. Caveolin-enriched membranes, isolated by either cationic silica or buoyant density methods, were explored by comparing large series of two-dimensional (2-D) maps and subsequent identification of over 100 protein spots by matrix-assisted laser desorption/ ionization (MALDI) peptide mass fingerprinting. Improved representation and identification of membrane proteins and valuable information on various post-translational modifications was achieved by the presented optimized procedures for solubilization, destaining and database searching/computing. Whereas the cationic silica purification yielded predominantly known endoplasmic reticulum residents, the cold-detergent method yielded a large number of known caveolae residents, including caveolin-1. Thus, a large part of this subproteome was established, including known (trans-)membrane, signal transduction and glycosyl phosphatidylinositol (GPI)-anchored proteins. Several predicted proteins from the human genome were isolated for the first time from biological samples, including SGRP58, SLP-2, C80RF2, and XRP-2. These findings and various optimized procedures can serve as a reference to study the differential composition of endothelial cell caveolae and rafts, known to be involved in pathologies like cancer and cardiovascular disease. [References: 41]
机译：人内皮细胞质膜具有两个类似脂质组成的亚结构域，即小窝和筏，它们都至关重要地参与了各种重要的细胞过程，如转胞吞作用，信号转导和胆固醇稳态。通过比较大系列的二维（2-D）图并随后通过基质辅助激光解吸/电离（MALDI）识别100多个蛋白质斑点，探索了通过阳离子二氧化硅或浮力密度法分离的富含卡夫林的膜肽质量指纹图谱。通过提出的用于增溶，脱色和数据库搜索/计算的优化程序，可以改善膜蛋白的表示和鉴定以及有关各种翻译后修饰的有价值的信息。阳离子二氧化硅的纯化主要产生已知的内质网居民，而冷洗涤剂方法产生了大量的已知小窝动物，包括小窝蛋白-1。因此，建立了该子蛋白质组的很大一部分，包括已知的（反式）膜，信号转导和糖基磷脂酰肌醇（GPI）锚定的蛋白质。首次从生物样品中分离出人类基因组中的几种预测蛋白，包括SGRP58，SLP-2，C80RF2和XRP-2。这些发现和各种优化的程序可以作为研究内皮细胞小窝和筏的不同成分的参考，这些小窝和筏被认为与癌症和心血管疾病等病理学有关。 [参考：41]

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