Fig. 2: Loss of aromaticity at residue 435 elicits structural and dynamic perturbations in the allosteric pocket.

a Chemical shift perturbation plots for Y435W (blue), Y435A (purple), and Y435S (orange) MKP5-CD. Colored bars represent ¹H¹⁵N combined chemical shift perturbations (Δδ), calculated for each mutant relative to WT MKP5-CD. Significant chemical shift perturbations were determined as those 1.5σ above the 10% trimmed mean of all data sets (gray horizontal line). Black vertical dashed lines denote resonances that exhibit line broadening, and gray vertical lines indicate residues that are unassigned. The site of mutation is noted by an asterisk above each plot. b Significant chemical shift perturbations are plotted onto the MKP5-CD structure (PDB: 2OUD) in blue (Y435W), purple (Y435A), and orange (Y435S) spheres. Line-broadened resonances are indicated by gray spheres, the site of mutation is denoted by a cyan sphere, and the side-chains of residues at the catalytic site (D377, C408, R414) are highlighted in red. c Representative snapshots from the ¹H¹⁵N HSQC spectral overlay of WT MKP5-CD (gray), Y435W (blue), Y435S (orange), and Y435A (purple) demonstrate that the Y435S and Y435A mutants exhibit similar chemical shift trajectories, suggesting a similar conformation populated by those mutants, while Y435W chemical shifts diverge from that of the serine and alanine mutants and often resemble WT. d Allosteric traffic maps of the MKP5-CD cartoon structure show the preferential communication routes in each state (WT, Y435W, Y435A, Y435S). e The main residues comprising each route are annotated.