Fig. 6

Kinase-mediated regulation of ECM dimer geometry in cancer-associated intracellular mutants. a Kinase domain structure showing the positions of relevant amino acids. b Fractions of FLImP measurements whose ranges of 69% confidence overlap with the ranges of DIII–DIII separations expected for dimers (0–15 nm) (pink) or oligomers (20–60 nm) (blue) collected from cells expressing the receptor mutants and/or under the conditions noted in the X-axis. The associated FLImP distributions are in Supplementary Fig. 18. c–e The FLImP distributions (grey) and the distributions compiled from the FLImP measurements whose 69% CIs overlap with DIII–DIII separations = 5 nm (green), 13 nm (yellow), or >20 nm (blue) collected from cells expressing wtEGFR, ΔC-EGFR, Δ698-EGFR, wtEGFR on cells treated with 1 μM erlotinib, wtEGFR on cells treated with 10 mM MβCD, L834R-EGFR, L680N-EGFR, Δ973-EGFR and T766M-EGFR. The insets show the fraction of separations consistent with each distance. Errors were assesed with bootstrap-resampling¹². f Free-energy surfaces as a function of the helical content of the αC helix and the distance from the reference active conformation (a contact map corresponding to the active extended A-loop conformation) as obtained from PTmetaD simulations of the wtEGFR and T766M-EGFR mutant. The central structures of the most populated clusters are shown (left and middle). The A-loop is coloured green, while the αC helix is shown in purple if it forms an α-helix and in pink if it forms a 3₁₀ helix. Salt-bridge interactions formed at the dimer interface in the last µs of the unbiased MD simulations of the symmetric dimers (right panel). The mean values and the standard deviations are calculated across monomers. The salt bridge was considered to be formed if the minimal distance between the side chains of residues in question was <4 Å. (More information on the calculations can be found in Supplementary Methods)