Supplementary Figure 6: Characterization of the Cdc14 Q106L and Cdc14 W108R mutant proteins.
From: A PxL motif promotes timely cell cycle substrate dephosphorylation by the Cdc14 phosphatase
(a), Cdc14 Q106L and Cdc14 W108R show uncompromised enzymatic activity. GFP-Cdc14, GFP-Cdc14 Q106L and GFP-Cdc14 W108R were purified and analyzed by SDS–PAGE followed by Coomassie blue staining. Velocity of p-NPP hydrolysis by GFP-Cdc14, GFP-Cdc14 Q106L and GFP-Cdc14 W108R was recorded as a function of the indicated substrate concentrations in three independent experiments. The means are shown; error bars indicate s.d. (b), GFP-Cdc14 Q106L and GFP-Cdc14 W108R fail to bind the Cbk1-derived PxL motif peptide. The means and s.d. from three independent microscale thermophoresis experiments to measure Cbk1-derived PxL motif peptide binding to the two Cdc14 variants are shown.
