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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Global mapping of rat plasma proteins with a native proteomic approach using nondenaturing micro 2DE and quantitative LC‐MS/MS
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Global mapping of rat plasma proteins with a native proteomic approach using nondenaturing micro 2DE and quantitative LC‐MS/MS

机译：大鼠血浆蛋白的全局蛋白质使用裸素蛋白质组学方法使用NondeNaturing Micro 2De和定量LC-MS / MS

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Plasma samples from adult male rats were separated by nondenaturing micro 2DE and a reference gel was selected, on which 136 CBB‐stained spots were numbered and subjected to in‐gel digestion and quantitative LC‐MS/MS. The analysis provided the assignment of 1–25 (average eight) non‐redundant proteins in each spot and totally 199 proteins were assigned in the 136 spots. About 40% of the proteins were detected in more than one spot and 15% in more than ten spots. We speculate this complexity arose from multiple causes, including protein heterogeneity, overlapping of protein locations and formation of protein complexes. Consequently, such results could not be appropriately presented as a conventional 2DE map, i.e. a list or a gel pattern with one or a few proteins annotated to each spot. Therefore, the LC‐MS/MS quantity data was used to reconstruct the gel distribution of each protein and a library containing 199 native protein maps was established for rat plasma. Since proteins that formed a complex would migrate together during the nondenaturing 2DE and thus show similar gel distributions, correlation analysis was attempted for similarity comparison between the maps. The protein pairs showing high correlation coefficients included some well‐known complexes, suggesting the promising application of native protein mapping for interaction analysis. With the importance of rat as the most commonly used laboratory animal in biomedical research, we expect this work would facilitate relevant studies by providing not only a reference library of rat plasma protein maps but a means for functional and interaction analysis.
机译：通过NondeNaturing Micro 2De分离成体阳性大鼠的等离子体样品，并选择参考凝胶，其中编号136cbb染色的斑点并进行凝胶分解和定量LC-MS / MS。分析提供了每次点的1-25（平均八个）非冗余蛋白的作用，并在136个斑点中分配了199999蛋白。在多个斑点中检测到约40％的蛋白质，超过十个斑点的15％。我们推测这种复杂性从多种原因产生，包括蛋白质异质性，蛋白质位置重叠和蛋白质复合物的形成。因此，这种结果不能被适当地呈现为常规的2DE图，即，具有一个或多种蛋白质的列表或凝胶模式，每个斑点。因此，LC-MS / MS量数据用于重建每种蛋白质的凝胶分布，并为大鼠等离子体建立含有199个天然蛋白质地图的文库。由于形成复合物的蛋白质将在NondEnaturing 2DE期间将它们迁移在一起，因此显示出类似的凝胶分布，因此尝试在地图之间进行相似性比较的相关性分析。显示高相关系数的蛋白质对包括一些众所周知的复合物，表明对天然蛋白质测绘进行相互作用分析的有望应用。随着大鼠作为生物医学研究中最常用的实验室动物的重要性，我们预计这项工作将通过提供大鼠血浆蛋白质地图的参考文库来促进相关研究，而是一种用于功能和相互作用分析的手段。

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