Fig. 3.

Impact of the gatekeeper substitution V561M on the kinase activity and overall structural changes of FGFR1 KD.

A. Left panel: comparison of the activity of FGFR1^WT and FGFR^V561M produced using the ADP-Glo Assay. Increasing concentrations of kinase were incubated for 60 min and the level of ADP produced analysed. Data were fit with a hyperbolic model (for presentation purposes only) using Graphpad prism. Each data point was produced in triplicate and the standard errors are indicated. Right panel: comparison of the enzyme efficiency (k_cat/K_m) of FGFR1^WT and FGFR^V561M. Parameters were generated through Michaelis–Menten kinetic experiments and analysed using Graphpad Prism software. See also Supplemental Tables S1 and S2.

B. The free energy surfaces of the activation of WT-FGFR and V561M-FGFR are shown as a function of the distance from the reference inactive A-loop conformation (CV1) and to the distance from the reference inactive A-loop conformation (CV2). The contour lines are drawn every 1 kcal/mol.

C. Ribbon representations of the proposed structures of FGFR1 WT and V561M coming from the full atomistic modelling molecular dynamics approach in the semi-inactive and active forms.