title: Solubilisation of the armadillo-repeat protein β-catenin using a zwitterionic detergent allows resolution of phosphorylated forms by 2DE.
abstract: β-catenin is a member of the armadillo repeat family of proteins and has important functions in cell-cell adhesion and Wnt signalling. Different protein species of β-catenin have been shown to exist in the cell and the relative proportions of these species are altered upon stimulation of cells with Wnt-3a (Gottardi and Gumbiner, 2004). In order to determine whether posttranslational modifications (PTMs) of β-catenin underlie these different protein species, we have used 2DE separation and immunoblotting with an antibody specific for β-catenin. High-resolution separation of differentially modified species of β-catenin in 2DE required the addition of ASB-16, a zwitterionic detergent that can solubilise integral membrane proteins. ASB-16 was also necessary for focussing of other armadillo repeat proteins, such as γ-catenin and p120-catenin. 2DE using ASB-16 allowed detection of a previously unreported phosphorylation site in the transcriptionally active form of β-catenin that binds to GST-Tcf in response to Wnt signalling.
authors: The Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Australia.
authors: Meredith J
authors: MJ
authors: Layton
authors: Nicole L
authors: NL
authors: Church
authors: Maree C
authors: MC
authors: Faux
authors: Hong
authors: H
authors: Ji
authors: Robert J A
authors: RJ
authors: Goode
authors: Eugene A
authors: EA
authors: Kapp
authors: Antony W
authors: AW
authors: Burgess
authors: Richard J
authors: RJ
authors: Simpson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000737:chemistry / Q000378:metabolism; D001622:Betaine / Q000031:analogs & derivatives* / Q000737:chemistry; D018938:Caco-2 Cells; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007525:Isoelectric Focusing; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D000073940:Transcription Factor 4; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D011994:Recombinant Proteins; C532096:TCF4 protein, human; D000073940:Transcription Factor 4; D014157:Transcription Factors; C561782:amidosulfobetaine-16; D051176:beta Catenin; D001622:Betaine