Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics
Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics
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DOI：
10.1038/nbt1005
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发表时间：
2004-09-01
影响因子：
46.9
通讯作者：
Mann, M
中科院分区：
文献类型：
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作者：
Blagoev, B;Ong, SE;Mann, M
为了研究早期生长因子刺激中基于磷酸酪氨酸的信号事件的全局动态，我们开发了一种质谱方法，该方法将时间变化转换为肽同位素丰度的差异。三个细胞群体的蛋白质组代谢编码不同的稳定同位素形式的精氨酸。每个群体刺激表皮生长因子不同的时间长度，酪氨酸磷酸化蛋白质和密切相关的粘合剂进行亲和纯化。含精氨酸的肽发生在三种形式，这是定量的，然后我们结合两个实验，以产生五点动态配置文件。我们确定了81个信号蛋白，包括几乎所有已知的表皮生长因子受体底物，31个新的效应器和它们在表皮生长因子刺激后激活的时间过程。全球激活配置文件提供了一个信息的角度对细胞信号，将是至关重要的建模信号网络的系统生物学方法。
To study the global dynamics of phosphotyrosine-based signaling events in early growth factor stimulation, we developed a mass spectrometric method that converts temporal changes to differences in peptide isotopic abundance. The proteomes of three cell populations were metabolically encoded with different stable isotopic forms of arginine. Each population was stimulated by epidermal growth factor for a different length of time, and tyrosine-phosphorylated proteins and closely associated binders were affinity purified. Arginine-containing peptides occurred in three forms, which were quantified; we then combined two experiments to generate five-point dynamic profiles. We identified 81 signaling proteins, including virtually all known epidermal growth factor receptor substrates, 31 novel effectors and the time course of their activation upon epidermal growth factor stimulation. Global activation profiles provide an informative perspective on cell signaling and will be crucial to modeling signaling networks in a systems biology approach.