Fig. 6. The Hinge, not the flap, mediates GSK2830371 binding.

A Surface plasmon resonance of PPM1D_Δflap and GSK2830371 (red represents raw data and black represents fitted kinetic parameters). B Relative differences in deuterium uptake between PPM1D_Δflap in the presence of GSK2830371 compared to PPM1D_Δflap in the absence of GSK2830371 as assessed by HDX-MS. Vertical tick marks between peptides on the X-axis are shown to indicate regions of missing coverage. The bars below the X-axis represent the N-terminus (purple), loop (brown), hinge (cyan), flap (light purple), and C-terminus (black) of the protein. C Activity of GSK2830371 in the FDP (top) and p38 MAPK (bottom) enzymatic assays with PPM1D_1–420 (black) or PPM1D_Δhinge (red). N = 2 independent samples runs. Data are presented as mean values ± SD. D EX1 analysis showing the change in deuterium uptake for peptide ₂₇₀ AVARALGDL ₂₇₈ located in the flap region in PPM1D_1–420 and PPM1D_Δhinge in the absence (left) and presence (right) of GSK2830731. Source data are provided in the Source Data file.