Supplementary Figure 8: The Cdc14 GETS dimer interface mutant.
From: A PxL motif promotes timely cell cycle substrate dephosphorylation by the Cdc14 phosphatase
(a), Additional illustration of the dimer interface within Cdc14, spanning both domains A and B and comprising extensive interactions between the two protomers. Residues at the dimer interface are displayed as sticks; a network of hydrogen bonds between the protomers is indicated. (b), Equal amounts of purified recombinant His6-Cdc14 and of the dimer interface mutant His6-Cdc14-GETS were analyzed by SDS–PAGE followed by Coomassie blue staining. (c), Analytical gel filtration profiles of Cdc14 and Cdc14-GETS using a Superdex 200 10/300 Increase column. The increased elution volume of Cdc14-GETS is suggestive of dimer disruption. (d), Velocity of p-NPP hydrolysis by 100 nM Cdc14 and Cdc14-GETS at the indicated substrate concentrations suggests that dimerization is required for full enzymatic activity of the phosphatase. The means and s.d. of three independent experiments are shown.
