Proteomics strategy for quantitative protein interaction profiling in cell extracts.
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.
Sharma
Kirti
K
Cell Signaling Group, Department of Molecular Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
Weber
Christoph
C
Bairlein
Michaela
M
Greff
Zoltán
Z
Kéri
György
G
Cox
Jürgen
J
Olsen
Jesper V
JV
Daub
Henrik
H
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*
D002457:Cell Extracts; D020543:Proteome