title: Use of selective extraction and fast chromatographic separation combined with electrophoretic methods for mapping of membrane proteins.
abstract: A model system for selective solubilization and fast separation of proteins from the rat liver membrane fraction and purified rat liver plasma membranes for their further proteomic analysis is presented. For selective solubilization, high-pH solutions and a concentrated urea solution, combined with different detergents, are used. After extraction, proteins are separated by anion-exchange chromatography or a combination of anion- and cation-exchange chromatography with convective interaction monolithic supports. This separation method enables fast and effective prefractionation of membrane proteins based on their hydrophobicity and charge prior to one-dimensional (1-D) and 2-D electrophoresis and mass spectrometry. By use of this sample preparation method, the less-abundant proteins can be detected and identified.
authors.affiliation: Proteomics Core, COBRE Center for Cancer Research Development, Rhode Island Hospital, The CORO Center, Providence, RI 02903, USA. Djuro.Josic@brown.edu
authors.forename: Djuro
authors.initials: D
authors.lastname: Josic
authors.forename: Mari Kino
authors.initials: MK
authors.lastname: Brown
authors.forename: Feilei
authors.initials: F
authors.lastname: Huang
authors.forename: Helen
authors.initials: H
authors.lastname: Callanan
authors.forename: Marijana
authors.initials: M
authors.lastname: Rucević
authors.forename: Alison
authors.initials: A
authors.lastname: Nicoletti
authors.forename: James
authors.initials: J
authors.lastname: Clifton
authors.forename: Douglas C
authors.initials: DC
authors.lastname: Hixson
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange; D015180:Electrophoresis, Gel, Two-Dimensional; D008099:Liver / Q000166:cytology; D008565:Membrane Proteins / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D051381:Rats
chemical_list: D008565:Membrane Proteins