Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., Dortmund, Germany;
Blood. 2014 Jan 30;123(5):e1-e10. doi: 10.1182/blood-2013-07-512384. Epub 2013 Dec 9.
One of the most important physiological platelet inhibitors is endothelium-derived prostacyclin which stimulates the platelet cyclic adenosine monophosphate/protein kinase A (cAMP/PKA)-signaling cascade and inhibits virtually all platelet-activating key mechanisms. Using quantitative mass spectrometry, we analyzed time-resolved phosphorylation patterns in human platelets after treatment with iloprost, a stable prostacyclin analog, for 0, 10, 30, and 60 seconds to characterize key mediators of platelet inhibition and activation in 3 independent biological replicates. We quantified over 2700 different phosphorylated peptides of which 360 were significantly regulated upon stimulation. This comprehensive and time-resolved analysis indicates that platelet inhibition is a multipronged process involving different kinases and phosphatases as well as many previously unanticipated proteins and pathways.
其中最重要的生理性血小板抑制剂之一是内皮衍生的前列环素，它刺激血小板环腺苷酸/蛋白激酶 A (cAMP/PKA)信号级联反应，几乎抑制所有血小板激活的关键机制。我们使用定量质谱法，在 3 个独立的生物学重复中，分析了人血小板在用稳定的前列环素类似物伊洛前列素处理 0、10、30 和 60 秒后的时间分辨磷酸化模式，以表征血小板抑制和激活的关键介质。我们定量分析了超过 2700 种不同的磷酸化肽，其中 360 种在刺激时显著调节。这种全面和时间分辨的分析表明，血小板抑制是一个多方面的过程，涉及不同的激酶和磷酸酶，以及许多以前未预料到的蛋白质和途径。
