title: Identifying nuclear protein-protein interactions using GFP affinity purification and SILAC-based quantitative mass spectrometry.
abstract: Many cellular proteins assemble into macromolecular protein complexes. Therefore, identifying protein-protein interactions (PPIs) is essential to gain insight into the function of proteins. Recently established quantitative mass spectrometry-based techniques have significantly improved the unbiased search for PPIs. In this chapter, we describe a single-step GFP affinity purification method combined with SILAC-based quantitative mass spectrometry that can be used to identify nuclear PPIs in mammalian cells.
authors: Department of Molecular Cancer Research, University Medical Center Utrecht, 85060, 3508 AB, Utrecht, The Netherlands.
authors: H Irem
authors: HI
authors: Baymaz
authors: Cornelia G
authors: CG
authors: Spruijt
authors: Michiel
authors: M
authors: Vermeulen
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D059748:Proteolysis; D014162:Transfection
chemical_list: D000596:Amino Acids; D009687:Nuclear Proteins; D010455:Peptides; D049452:Green Fluorescent Proteins