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Technical limitations have prevented proteomic analyses ofevents occurring less than 30 s after signal initiation. Wedeveloped an automated, continuous quench-flow systemallowing quantitative proteomic assessment of very early cellularsignaling events (qPACE) with a time resolution of 1 s. Usingthis technique, we determined that autophosphorylation of theepidermal growth factor receptor occurs within 1 s after ligandstimulation and is followed rapidly by phosphorylation of thedownstream signaling intermediates Src homologous andcollagen-like protein and phospholipase C gamma 1

Quantitative proteomic assessment of very early cellular signaling events: qPACE