Brief Communication abstract
Nature Methods 6, 741 - 744 (2009)
Published online: 13 September 2009 | doi:10.1038/nmeth.1373
Proteomics strategy for quantitative protein interaction profiling in cell extracts
Kirti Sharma1,6,7, Christoph Weber1,7, Michaela Bairlein2,7, Zoltán Greff3, György Kéri3,4, Jürgen Cox5, Jesper V Olsen5,6 & Henrik Daub1
We report a proteomics strategy to both identify and quantify cellular target protein interactions with externally introduced ligands. We determined dissociation constants for target proteins interacting with the ligand of interest by combining quantitative mass spectrometry with a defined set of affinity purification experiments. We demonstrate the general utility of this methodology in interaction studies involving small-molecule kinase inhibitors, a tyrosine-phosphorylated peptide and an antibody as affinity ligands.
- Cell Signaling Group, Department of Molecular Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
- Department of Molecular Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
- Vichem Chemie Ltd., Budapest, Hungary.
- Pathobiochemistry Research Group of the Hungarian Academy of Science, Semmelweis University, Budapest, Hungary.
- Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
- Present addresses: ITC Research and Development Centre, Survey Nos 230 and 243, Turkapally Village, Hyderabad, India (K.S.) and Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark (J.V.O.).
- These authors contributed equally to this work.
Correspondence to: Henrik Daub1 e-mail: daub@biochem.mpg.de
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