Quantitative proteomic assessment of very early cellular signaling events.
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.
Dengjel
Joern
J
Center of Experimental BioInformatics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Campusvej 55, DK-5230, Odense, Denmark.
Akimov
Vyacheslav
V
Olsen
Jesper V
JV
Bunkenborg
Jakob
J
Mann
Matthias
M
Blagoev
Blagoy
B
Andersen
Jens S
JS
D066246:ErbB Receptors / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D008657:Metabolic Clearance Rate; D008856:Microscopy, Fluorescence / Q000379:methods*; D010766:Phosphorylation; D020543:Proteome / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013997:Time Factors
D020543:Proteome; D066246:ErbB Receptors