FIGURE 7.

EF2K binds to βTRCP1 and βTRCP2. A, HEK cells were co-transfected with GFP-EF2K and one of a panel of GST-tagged F-box proteins. Cell lysates were incubated with glutathione beads, and binding was assessed in inputs (upper panel) and eluates (lower panel) by performing SDS-PAGE and immunoblot for GFP and GST. * indicates GST-Fbw7a, which migrates at a similar but distinct molecular weight as GFP-EF2K by SDS-PAGE. B, HEK cells were co-transfected with GST-βTRCP1, and either wild-type GFP-EF2K, or S440/S444 AA, or Y442F mutant forms and pull-downs were performed as above. Binding was assessed in inputs (upper panel) and eluates (lower panel) by performing SDS-PAGE and immunoblot for GFP and GST.