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. Author manuscript; available in PMC: 2010 Nov 25.
Published in final edited form as: Mol Cell. 2009 Nov 25;36(4):547–559. doi: 10.1016/j.molcel.2009.09.034

Figure 2. CK2α Interacts with and Phosphorylates α-catenin at S641.

Figure 2

(B-E) Immunoblotting analyses were performed with the indicated antibodies.

(A) α-catenin immunoprecipitated from EGF-stimulated A431 cells was analyzed with mass spectrometry. Mass spectrometric analysis of a tryptic fragment 634-TPEELDDSDFETEDFDVR-651 indicates S641 was phosphorylated. The m/z difference between y-11 and y-10 matched with phospho-Ser.

(B) In vitro kinase assays were performed with purified bacterially expressed WT His-α-catenin or His-α-catenin S641A with or without CK2α.

(C) pRc/CMV2-HA-CK2α was co-transfected with pEGFP N1-FLAG-tagged WT α-catenin or α-catenin S641A mutant into 293T cells. Immunoblotting of immunoprecipitated FLAG-tagged α-catenins with the anti–α-catenin Ser641 antibody was performed.

(D) The subcellular fractionation was prepared from A431 cells treated with EGF (100 ng/ml) for 30 min.

(E) A431 cells expressing FLAG-tagged WT α-catenin were treated with or without EGF (100 ng/ml) for 1 or 3 hr. CK2α was immunoprecipitated.

(F) A431 cells with or without expression of CK2α shRNA or a control shRNA were treated with or without EGF (100 ng/ml) for 30 min.