Solubilisation of the armadillo-repeat protein beta-catenin using a zwitterionic detergent allows resolution of phosphorylated forms by 2DE
- Author(s)
- Layton, Meredith J; Church, Nicole L; Faux, Maree C; Ji, Hong; Goode, Robert JA; Kapp, Eugene A; Burgess, Antony W; Simpson, Richard J;
- Details
- Publication Year 2012-07,Volume 33,Issue #12,Page 1804-1813
- Journal Title
- Electrophoresis
- Publication Type
- Journal Article
- 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.
- Publisher
- WILEY
- Keywords
- Amidosulfobetaine;2Dimensional gel electrophoresis;Phosphorylation;Posttranslational modification;Wnt signalling
- Publisher's Version
- https://doi.org/10.1002/elps.201100671
- Terms of Use/Rights Notice
- © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Creation Date: 2014-02-27 01:08:06