Exploring the platelet proteome via combinatorial, hexapeptide ligand libraries
Exploring the platelet proteome via combinatorial, hexapeptide ligand libraries
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DOI：
10.1021/pr0703371
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发表时间：
2007-11-01
影响因子：
4.4
通讯作者：
Righetti, Pier Giorgio
中科院分区：
文献类型：
--
作者：
Guerrier, Luc;Claverol, Stephane;Righetti, Pier Giorgio
组合配体库，由数百万个不同的六肽诱饵，能够捕获和集中的“低丰度”的蛋白质组，同时大幅削减浓度的最丰富的物种，已被应用于探索可溶性血小板蛋白质组。未经处理和库处理的血小板的质谱分析导致鉴定出435种独特的基因产物。其中147个条目（占总数的35%）在以前报道的血小板蛋白质组研究中的> 1100种蛋白质列表中未被描述。此外，从二维电泳分析切除点的分析允许添加其他57种蛋白质，这些蛋白质在LC-MS分析中没有发现，其中33种在蛋白质组学研究中没有描述过，使新基因产物的总数达到180。因此，本数据增加了不可忽略的数量的物种，以继续血小板的蛋白质组资产的“制图”，鉴于完成映射程序，以更深入地了解这种血细胞的生理学和病理学。因为捕获过程是在生理条件下通过利用天然蛋白质构型结合组合文库进行的，所以所述技术不适于捕获高度疏水的蛋白质，其需要与我们的工作程序不相容的强变性和溶解条件。因此，我们的列表报告了基本上亲水的蛋白质，具有负的GRAVY指数。
A combinatorial ligand library, composed of millions of diverse hexapeptide baits, able to capture and concentrate the "low-abundance" proteome while drastically cutting the concentration of the most abundant species, has been applied to the exploration of the soluble platelet proteome. Mass spectrometry analysis of untreated and library-treated platelets has resulted in the identification of 435 unique gene products. Of those, 147 entries (35% of the total) have not been described among the list of > 1100 proteins in proteomic platelet investigations reported before. In addition, the analysis of excised spots from two-dimensional electrophoresis analysis allowed 57 other proteins to be added that were not found in LC-MS analysis, 33 of them not described before in proteomics studies, bringing the total number of new gene products to 180. Thus, the present data add a non-negligible number of species for continuing the "cartography" of the proteomic asset of platelets, in view of completing the mapping procedure for a deeper understanding of the physiology and pathology of this blood cell. Because the capturing process is performed under physiological conditions, by exploiting, for binding to the combinatorial library, the native protein configuration, the described technique is not adapted to capture highly hydrophobic proteins, which need strong denaturing and solubilizing conditions that are incompatible with our working procedure. Thus, our list reports essentially hydrophilic proteins, with negative GRAVY indexes.