title: Red wine proteins: two dimensional (2-D) electrophoresis and mass spectrometry analysis.
abstract: The aim of the present study was to optimize protein extraction from red wine (cv. Cabernet) in order to obtain a separation by two-dimensional electrophoresis (2-DE) compatible with mass spectrometry identification. Proteins were denatured by sodium dodecyl-sulphate (SDS) and precipitated as potassium salts. The potassium-DS (KDS) protein complexes obtained were treated with different solutions in order to remove the detergent. Proteins were solubilized with different buffers and separated by different electrophoretic approaches [native, urea, acid urea PAGEs and isoelectric focusing (IEF)] as the first-dimension (1-DE). The best 2D separation was achieved by using 10% saccharose in the DS removal step, and 6-cyclohexylhexyl β-d-maltoside detergent in the solubilisation buffer combined with the IEF approach. Several well focalized protein spots were obtained and analyzed through mass-spectrometry.
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Mainente
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Zoccatelli
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Marilinda
authors.initials: M
authors.lastname: Lorenzini
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Cecconi
authors.affiliation: Department of Agronomy Food Natural Resources Animals and Environments (DAFNAE), University of Padova, Viale dell'Università, 35020 Legnaro, Padova, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Vincenzi
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Corrado
authors.initials: C
authors.lastname: Rizzi
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy. Electronic address: barbara.simonato@univr.it.
authors.forename: Barbara
authors.initials: B
authors.lastname: Simonato
mesh_terms: D001203:Ascomycota / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000032:analysis; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D010940:Plant Proteins / Q000032:analysis*; D012967:Sodium Dodecyl Sulfate; D027843:Vitis / Q000737:chemistry*; D014920:Wine / Q000032:analysis* / Q000592:standards
chemical_list: D005656:Fungal Proteins; D010940:Plant Proteins; D012967:Sodium Dodecyl Sulfate
keywords: Mass spectrometry; Red wine proteins; Two-dimensional electrophoresis; Wine proteins precipitation