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. 2009 Oct;6(10):741-4.
doi: 10.1038/nmeth.1373. Epub 2009 Sep 13.

Proteomics strategy for quantitative protein interaction profiling in cell extracts

Kirti Sharma  1 , Christoph Weber, Michaela Bairlein, Zoltán Greff, György Kéri, Jürgen Cox, Jesper V Olsen, Henrik Daub
Affiliations

Proteomics strategy for quantitative protein interaction profiling in cell extracts

Kirti Sharma et al. Nat Methods. 2009 Oct.

Abstract

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.

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