title: Using in-solution digestion, peptide fractionation, and a Q exactive mass spectrometer to analyze the proteome of clathrin-coated vesicles.
abstract: The characterization of clathrin-coated vesicles (CCVs), including the effects of genetic or biochemical manipulations on their composition, can be studied by mass spectrometry analysis of HeLa cell fractions enriched for CCVs. This protocol describes the preparation of samples by in-solution proteolytic digest and subsequent peptide fractionation, followed by analysis in a Q Exactive mass spectrometer.
authors.affiliation: Department of Proteomics and Signal Transduction, Max Plank Institute of Biochemistry, 82152 Martinsried, Germany;
authors.forename: Georg H H
authors.initials: GH
authors.lastname: Borner
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, United Kingdom.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Fielding
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D022163:Clathrin-Coated Vesicles / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D059748:Proteolysis; D020543:Proteome / Q000032:analysis*
chemical_list: D020543:Proteome