In Vivo Identification of Plant Protein Complexes Using IP-MS/MS.
Individual proteins often function as part of a protein complex. The identification of interacting proteins is therefore vital to understand the biological role and function of the studied protein. Here we describe a method for the in vivo identification of nuclear, cytoplasmic, and membrane-associated protein complexes from plant tissues using a strategy of immunoprecipitation followed by tandem mass spectrometry. By performing quantitative mass spectrometry measurements on biological triplicates, relative abundance of proteins in GFP-tagged complexes compared to background controls can be statistically evaluated to identify high-confidence interactors. We detail the entire workflow of this approach.
['Wendrich|Jos R|JR|', 'Boeren|Sjef|S|', 'Möller|Barbara K|BK|', 'Weijers|Dolf|D|', 'De Rybel|Bert|B|']
D002462:Cell Membrane D002467:Cell Nucleus D003593:Cytoplasm D047468:Immunoprecipitation D008565:Membrane Proteins D010940:Plant Proteins D053719:Tandem Mass Spectrometry
Immunoprecipitation Tandem mass spectrometry Arabidopsis GFP Protein complex identification Protein–protein interaction