TY - JOUR
T1 - Global, in vivo, and site-specific phosphorylation dynamics in signaling networks
AU - Olsen, Jesper Velgaard
AU - Blagoev, Blagoy
AU - Gnad, Florian
AU - Macek, Boris
AU - Kumar, Chanchal
AU - Mortensen, Peter
AU - Mann, Matthias
N1 - Keywords: Binding Sites; Databases, Factual; Epidermal Growth Factor; Hela Cells; Humans; Kinetics; Mass Spectrometry; Neoplasm Proteins; Peptides; Phosphorylation; Phosphotyrosine; Protein Binding; Protein Processing, Post-Translational; Proteome; Signal Transduction
PY - 2006
Y1 - 2006
N2 - Cell signaling mechanisms often transmit information via posttranslational protein modifications, most importantly reversible protein phosphorylation. Here we develop and apply a general mass spectrometric technology for identification and quantitation of phosphorylation sites as a function of stimulus, time, and subcellular location. We have detected 6,600 phosphorylation sites on 2,244 proteins and have determined their temporal dynamics after stimulating HeLa cells with epidermal growth factor (EGF) and recorded them in the Phosida database. Fourteen percent of phosphorylation sites are modulated at least 2-fold by EGF, and these were classified by their temporal profiles. Surprisingly, a majority of proteins contain multiple phosphorylation sites showing different kinetics, suggesting that they serve as platforms for integrating signals. In addition to protein kinase cascades, the targets of reversible phosphorylation include ubiquitin ligases, guanine nucleotide exchange factors, and at least 46 different transcriptional regulators. The dynamic phosphoproteome provides a missing link in a global, integrative view of cellular regulation.
AB - Cell signaling mechanisms often transmit information via posttranslational protein modifications, most importantly reversible protein phosphorylation. Here we develop and apply a general mass spectrometric technology for identification and quantitation of phosphorylation sites as a function of stimulus, time, and subcellular location. We have detected 6,600 phosphorylation sites on 2,244 proteins and have determined their temporal dynamics after stimulating HeLa cells with epidermal growth factor (EGF) and recorded them in the Phosida database. Fourteen percent of phosphorylation sites are modulated at least 2-fold by EGF, and these were classified by their temporal profiles. Surprisingly, a majority of proteins contain multiple phosphorylation sites showing different kinetics, suggesting that they serve as platforms for integrating signals. In addition to protein kinase cascades, the targets of reversible phosphorylation include ubiquitin ligases, guanine nucleotide exchange factors, and at least 46 different transcriptional regulators. The dynamic phosphoproteome provides a missing link in a global, integrative view of cellular regulation.
U2 - 10.1016/j.cell.2006.09.026
DO - 10.1016/j.cell.2006.09.026
M3 - Journal article
C2 - 17081983
SN - 0092-8674
VL - 127
SP - 635
EP - 648
JO - Cell
JF - Cell
IS - 3
ER -