title: Efficient extraction of nucleolar proteins for interactome analyses.
abstract: The efficient extraction of proteins from purified cellular organelles is critical for in vitro analyses, including identification of protein complex members by affinity purification-based quantitative proteomic approaches. When applied to purified nucleoli, classic nuclear protein extraction methods inefficiently and selectively release only approximately 50% of proteins. Here, we present a method that can extract up to 90% of nucleolar proteins, and apply it in a quantitative interactomic approach to identify nucleolar interaction partners for a mammalian protein phosphatase.
authors.affiliation: Department of Cellular & Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
authors.forename: Delphine
authors.initials: D
authors.lastname: Chamousset
authors.forename: Sarah
authors.initials: S
authors.lastname: Mamane
authors.forename: François-Michel
authors.initials: FM
authors.lastname: Boisvert
authors.forename: Laura
authors.initials: L
authors.lastname: Trinkle-Mulcahy
mesh_terms: D002247:Carbon Isotopes; D002466:Cell Nucleolus / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D007553:Isotope Labeling; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D014357:Trypsin / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010749:Phosphoprotein Phosphatases; D014357:Trypsin