Figure 4. A peptide from photoreceptor cilium actin regulator (PCARE) achieves paralog selectivity by stabilizing an ENAH EVH1 domain-specific conformation.
(A) Superposition of ENAH EVH1 domains bound to PCARE or FP4 peptide (PDB 1EVH). The black arrow highlights a 3 Å shift in a loop that forms part of the binding pocket. Insets show residues that differ between ENAH and VASP or EVL near this loop. (B) Lowest dTERMen energy obtained when swapping 0–6 residues from ENAH into EVL, when modeled on the structure of ENAH EVH1 bound to PCARE. * indicates the mutation was added based on manual inspection. (C) ENAH EVH1 domain bound to a peptide from PCARE, with residues that were swapped into the EVL EVH1 domain to rescue affinity marked as purple spheres. On the right are binding curves for WT EVL EVH1 domain and EVLswapped EVH1 domain binding to PCARE B. Error reported as the standard deviation of two replicates.
Figure 4—figure supplement 1. Fast protein liquid chromatography (FPLC) curves for EVL mutants.
