Supplementary Figure 1: Biochemical characterization of the PxL motif.
From: A PxL motif promotes timely cell cycle substrate dephosphorylation by the Cdc14 phosphatase
(a), Recombinant Cdc14 preparations used for the phage display screen (His-Cdc14), peptide array (HA-Cdc14) and microscale thermophoresis experiments (GFP-Cdc14) were analyzed by SDS–PAGE and stained with Coomassie blue. A size marker is included. (b), Peptides recovered in the phage display screen are enriched in known Cdc14 substrates and Cdc14 interactors12,19 (hypergeometric test). (c), Mutational Cbk1 peptide array to probe the contribution of individual positions to Cdc14 binding. As in Fig. 1c, but a different membrane solvation was used (bottom), as indicated. A picture of the peptides stained with Ponceau S is also shown (top), as well as a control using the anti-HA antibody only (middle). (d), Microscale thermophoresis profiles of additional variant Cbk1-derived PxL motif peptides binding to GFP-Cdc14. Shown are the means and s.d. from two (F82G, V85G and Δ91–97) or three (L88G) independent experiments.
