Kinase-Selective Enrichment Enables Quantitative Phosphoproteomics of the Kinome across the Cell Cycle

Henrik Daub, Jesper V. Olsen, Michaela Bairlein, Florian Gnad, Felix S. Oppermann, Roman Körner, Zoltán Greff, György Kéri, Olaf Stemmann, Matthias Mann

Research output: Contribution to journalArticle

445 Citations (Scopus)

Abstract

Protein kinases are pivotal regulators of cell signaling that modulate each other's functions and activities through site-specific phosphorylation events. These key regulatory modifications have not been studied comprehensively, because low cellular abundance of kinases has resulted in their underrepresentation in previous phosphoproteome studies. Here, we combine kinase-selective affinity purification with quantitative mass spectrometry to analyze the cell-cycle regulation of protein kinases. This proteomics approach enabled us to quantify 219 protein kinases from S and M phase-arrested human cancer cells. We identified more than 1000 phosphorylation sites on protein kinases. Intriguingly, half of all kinase phosphopeptides were upregulated in mitosis. Our data reveal numerous unknown M phase-induced phosphorylation sites on kinases with established mitotic functions. We also find potential phosphorylation networks involving many protein kinases not previously implicated in mitotic progression. These results provide a vastly extended knowledge base for functional studies on kinases and their regulation through site-specific phosphorylation.

Original languageEnglish
Pages (from-to)438-448
Number of pages11
JournalMolecular Cell
Volume31
Issue number3
DOIs
Publication statusPublished - Aug 8 2008

Keywords

  • CELLCYCLE
  • SIGNALING
  • SYSBIO

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Daub, H., Olsen, J. V., Bairlein, M., Gnad, F., Oppermann, F. S., Körner, R., Greff, Z., Kéri, G., Stemmann, O., & Mann, M. (2008). Kinase-Selective Enrichment Enables Quantitative Phosphoproteomics of the Kinome across the Cell Cycle. Molecular Cell, 31(3), 438-448. https://doi.org/10.1016/j.molcel.2008.07.007