Protein interaction screening by quantitative immunoprecipitation combined with knockdown (QUICK).
Present screening methods for protein-protein interactions (PPIs) rely on the overexpression of artificial fusion proteins, making it difficult to assess in vivo relevance. Here we combine stable isotope labeling with amino acids in cell culture (SILAC), RNA interference (RNAi), coimmunoprecipitation and quantitative mass-spectrometry analysis to detect cellular interaction partners of endogenous proteins in mammalian cells with very high confidence. We used this screen to identify interaction partners of beta-catenin and Cbl.
Selbach
Matthias
M
Max Planck Institute of Biochemistry, Department of Proteomics and Signal Transduction, Am Klopferspitz 18, D-82152 Martinsried, Germany.
Mann
Matthias
M
D020868:Gene Silencing; D007118:Immunoassay / Q000379:methods* / Q000639:trends*; D047468:Immunoprecipitation / Q000379:methods* / Q000639:trends*; D013058:Mass Spectrometry / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods* / Q000639:trends*