Quantitative proteomic assessment of very early cellular signaling events
Quantitative proteomic assessment of very early cellular signaling events
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DOI：
10.1038/nbt1301
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发表时间：
2007-05-01
影响因子：
46.9
通讯作者：
Andersen, Jens S.
中科院分区：
文献类型：
--
作者：
Dengjel, Joern;Akimov, Vyacheslav;Andersen, Jens S.
技术限制阻碍了对信号启动后 30 秒内发生的事件进行蛋白质组学分析。我们开发了一种自动化、连续的猝灭流系统，可以以 1 秒的时间分辨率对非常早期的细胞信号事件 (qPACE) 进行定量蛋白质组学评估。利用这种技术，我们确定表皮生长因子受体的自身磷酸化发生在配体刺激后 1 秒内，随后下游信号中间体 Src 同源蛋白、胶原蛋白样蛋白和磷脂酶 C gamma 1 迅速磷酸化。
Technical limitations have prevented proteomic analyses of events occurring less than 30 s after signal initiation. We developed an automated, continuous quench-flow system allowing quantitative proteomic assessment of very early cellular signaling events (qPACE) with a time resolution of 1 s. Using this technique, we determined that autophosphorylation of the epidermal growth factor receptor occurs within 1 s after ligand stimulation and is followed rapidly by phosphorylation of the downstream signaling intermediates Src homologous and collagen-like protein and phospholipase C gamma 1.