title: Proteomics strategy for quantitative protein interaction profiling in cell extracts.
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.
authors.affiliation: Cell Signaling Group, Department of Molecular Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Kirti
authors.initials: K
authors.lastname: Sharma
authors.forename: Christoph
authors.initials: C
authors.lastname: Weber
authors.forename: Michaela
authors.initials: M
authors.lastname: Bairlein
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Greff
authors.forename: György
authors.initials: G
authors.lastname: Kéri
authors.forename: Jürgen
authors.initials: J
authors.lastname: Cox
authors.forename: Jesper V
authors.initials: JV
authors.lastname: Olsen
authors.forename: Henrik
authors.initials: H
authors.lastname: Daub
mesh_terms: D002457:Cell Extracts / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
chemical_list: D002457:Cell Extracts; D020543:Proteome