Piersma Sander R, Broxterman Henk J, Kapci Muhammed, de Haas Richard R, Hoekman Klaas, Verheul Henk M W, Jiménez Connie R
OncoProteomics Laboratory, Dept. of Medical Oncology, VUmc-Cancer Center Amsterdam, VU University Medical Center Amsterdam, Amsterdam, The Netherlands.
J Proteomics. 2009 Feb 15;72(1):91-109. doi: 10.1016/j.jprot.2008.10.009. Epub 2008 Nov 8.
Upon stimulation, platelets release the soluble content of their cytoplasmic granules along with microparticles. This sub-proteome is of interest since many of its constituents are associated with coagulation, (tumor) angiogenesis, cell growth and adhesion. Previously, differential - antibody-based - serum analysis has yielded information on the proteins released from platelets upon stimulation. A promising alternative strategy is formed by identifying the proteins released by freshly isolated platelets from blood using proteomics. Here we report on the analysis of the thrombin receptor activating peptide (TRAP)-induced releasate from 3 different volunteers using high resolution, high mass accuracy hybrid LTQ-FT mass spectrometry in a GeLC-MS/MS workflow. We obtained an activated platelet releasate proteome comprising a total of 716 identified proteins with 225 proteins present in the releasate of 3/3 volunteers. This core dataset is characterized by gene ontology mining and signal peptide analysis. Meta-analysis of our dataset and two published datasets of platelet a-granules and microparticles reveals that 55% of our platelet releasate proteins can be annotated using these previous platelet subproteome data, of the remaining releasate proteome 5% overlaps with a published platelet secretome while 40% of our data consists of novel releasate proteins. This high-accuracy activated platelet releasate proteome represents the largest and most comprehensive analysis to date. This approach offers unique possibilities to analyse the role of platelet-secreted proteins in physiology and in diseases such as atherosclerosis and cancer.
受到刺激后，血小板会释放其细胞质颗粒的可溶性成分以及微粒。这个亚蛋白质组备受关注，因为其许多成分都与凝血、（肿瘤）血管生成、细胞生长和黏附有关。此前，基于差异抗体的血清分析已得出关于刺激后血小板释放的蛋白质的信息。一种有前景的替代策略是通过蛋白质组学鉴定从血液中新鲜分离的血小板释放的蛋白质。在此，我们报告了在GeLC-MS/MS工作流程中使用高分辨率、高质量精度的混合LTQ-FT质谱对3名不同志愿者的凝血酶受体激活肽（TRAP）诱导的释放物进行的分析。我们获得了一个活化血小板释放蛋白质组，共鉴定出716种蛋白质，其中225种蛋白质存在于3名志愿者全部的释放物中。这个核心数据集通过基因本体挖掘和信号肽分析进行了表征。对我们的数据集与两个已发表的血小板α颗粒和微粒数据集的荟萃分析表明，我们的血小板释放物蛋白质中有55%可以使用这些先前的血小板亚蛋白质组数据进行注释，其余释放物蛋白质组中有5%与已发表的血小板分泌蛋白质组重叠，而我们的数据中有40%由新的释放物蛋白质组成。这个高精度的活化血小板释放蛋白质组代表了迄今为止最大且最全面的分析。这种方法为分析血小板分泌蛋白在生理以及动脉粥样硬化和癌症等疾病中的作用提供了独特的可能性。
