Comparison of human platelet membrane-cytoskeletal proteins with the plasma proteome: Towards understanding the platelet-plasma nexus
Comparison of human platelet membrane-cytoskeletal proteins with the plasma proteome: Towards understanding the platelet-plasma nexus
复制标题
DOI：
10.1002/prca.200780067
复制
发表时间：
2008-01-01
影响因子：
2
通讯作者：
Simpson, Richard J.
中科院分区：
文献类型：
--
作者：
Greening, David W.;Glenister, Kristen M.;Simpson, Richard J.
血小板对于维持血管完整性至关重要。鉴于血小板的无核性质，其蛋白质组的定义是必不可少的理解血小板的病理生理。我们在这里描述了对纯化的、正常的、未活化的人血小板的血小板膜/细胞骨架亚蛋白质组进行的详细的基于MS的蛋白质组学分析。与之前的血小板蛋白质组学纯化策略相比，本研究中使用血沉棕黄层方法分离和纯化最低限度活化的血小板，显著降低了白细胞（0.02 +/- 0.007 x 10（6）/L）和红细胞（0.21 +/- 0.02%）的污染物。使用1%的错误发现率，203个蛋白质进行了鉴定，并就其亚细胞定位，生物学功能和细胞过程进行了表征。其中16种在以前的人类血小板蛋白质组研究中尚未鉴定。作为了解动态血小板-血浆蛋白组成关系的第一种方法，我们重新分析了整个HUPO血浆蛋白质组项目数据集（鉴定了647种血浆蛋白质），并将这些数据与我们的血小板蛋白质组数据集进行了比较。进一步分析了共同鉴定的蛋白质（41）在这两种蛋白质组中的相对丰度（指数修正的蛋白质丰度指数）和功能富集，以及它们与血小板转录组的相关性。血小板膜/细胞骨架和血浆蛋白质组参考数据集（包括处理和未处理的MS/MS光谱）均可公开访问（http：//www.ludwig.edu.au/archive/）。
Platelets are essential for maintaining vascular integrity. Given the anucleate nature of platelets, definition of their proteome is essential for understanding platelet pathophysiology. We describe here a detailed MS-based proteomic analysis of the platelet membrane/cytoskeletal sub-proteome from purified, normal, non-activated human platelets. In contrast to previous platelet proteomic purification strategies, the buffy-coat method was utilized in this study to isolate and purify minimally activated platelets, yielding significantly reduced contaminants for leukocytes (0.02 +/- 0.007 x 10(6)/L) and erythrocytes (0.21 +/- 0.02%). Using a false discovery rate of 1%, 203 proteins were identified and characterized with respect to their subcellular localization, biological function, and cellular processes. of these, 16 have not been identified in previous human platelet proteome studies. As a first approach towards understanding the dynamic platelet-plasma protein composition nexus, we re-analysed the entire HUPO plasma proteome project dataset (647 plasma proteins identified) and compared these data with our platelet proteome dataset. Co-identified proteins (41) were further analysed with respect to their relative abundances (exponentially modified protein abundance index) and functional enrichment in these two proteomes, as well as their correlation with the platelet transcriptome. Both platelet membrane/cytoskeletal and plasma proteome reference datasets, comprising both processed and unprocessed MS/MS spectra, are publicly accessible (http://www.ludwig.edu.au/archive/).