Comparative analysis of platelet-derived microparticles reveals differences in their amount and proteome depending on the platelet stimulus
Comparative analysis of platelet-derived microparticles reveals differences in their amount and proteome depending on the platelet stimulus
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DOI：
10.1016/j.jprot.2012.02.030
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发表时间：
2012-12-05
影响因子：
3.3
通讯作者：
Garcia, Angel
中科院分区：
文献类型：
--
作者：
Shai, Ela;Rosa, Isaac;Garcia, Angel
血小板衍生微粒（PMP）在许多疾病（例如心血管疾病和癌症）中升高。在本研究中，我们进行了高分辨率的2-DE为基础的蛋白质组学分析的PMP起源后，血小板活化与不同的刺激。流式细胞术分析显示，与0.5 U/mL凝血酶相比，当血小板被剪切激活时（1800 s（-1）），微粒的平均数量更高。关于蛋白质组学分析，发现30个蛋白质特征在pI 4-7范围内在剪切和凝血酶组之间差异调节，其中28个通过MS成功鉴定，对应于26个开放阅读框。信号蛋白构成了主要的功能群，包括膜受体和衔接子。不可逆途径分析（IPA）软件显示，26个差异调节的独特蛋白质中有21个是与细胞组装和组织以及细胞形态相关的共同网络的一部分。蛋白质印迹分析证实，Dok-2和整合素α 6在PMP中受到不同的调节，具体取决于血小板刺激。有趣的是，这两种蛋白质都参与调节血管生成的机制，因此可能是PMP调节内皮细胞功能的一部分。总之，这项研究表明，血小板脱落微粒的数量不同，并与不同的蛋白质组取决于stimulation.This文章的一部分，题为：综合组学特刊。(C)2012爱思唯尔有限公司版权所有。
Platelet-derived microparticles (PMP) are elevated in a number of disorders (e.g. cardiovascular disease and cancer). In the present study, we have carried out a high-resolution 2-DE-based proteomic analysis of PMP originated following platelet activation with different stimulus. Flow cytometric analysis revealed a higher average number of microparticles when platelets are activated with shear (1800 s(-1)) compared to 0.5 U/mL thrombin. Regarding the proteomic analysis, 30 protein features were found to be differentially regulated between shear and thrombin groups in the pI 4-7 range, from which 28 were successfully identified by MS, corresponding to 26 open-reading frames. Signaling proteins constituted the major functional group, including membrane receptors, and adapters. Ingenuity Pathways Analysis (IPA) software revealed that 21 of the 26 differentially regulated unique proteins identified are part of a common network related to cell assembly and organization and cell morphology. Western blotting analyses confirmed that Dok-2 and the integrin alpha 6 are differentially regulated in PMP depending on the platelet stimulus. Interestingly, both proteins participate in mechanisms regulating angiogenesis so could be part of PMP regulation of endothelial cells function. In conclusion, this study shows that platelets shed microparticles in different amounts and with a different proteome depending on the stimulus.This article is part of a Special Issue entitled: Integrated omics. (C) 2012 Elsevier B.V. All rights reserved.