wildtype dimerL834R dimer

deletion dimer

peptide docked monomer

RMSD from inactive (Å )

RM

SD

fro

m a

ctiv

e (Å

)

The novel asymmetric dimer interface with the activating EGFR in brown and the active EGFR in sky blue. The dimer interfaces are shown in lime and orange respectively, with the A-loop in red, C-loop in blue and the N-loop in green.

C-helix

A-loop

Trajectory of the umbrella sampling from inactive to active EGFR. The A-loop and C-helix are in red and blue, with the initial conformation of both shown in green.

R812 D813

L834

K836

E738

D737

K721

L679

A743

R865H846

E848

K851

Examination of the stabilizing inactivating network breaking as EGFR becomes more active. The inactive bonds (residues in white, atoms in green) must break before moving and the active bonds (residues in orange, atoms in purple) must form as EGFR activates.

D813

L834

K836

D738

K721

Enhanced view of the inactivation of key catalytic residues. The catalytic aspartate (D813) is bonded with L834 and must be broken before catalysis can occur. While a conserved salt bridge (K721-D738) is broken with D738 bonded to K836. K836 must flip while D738 rotates to bond to K721 for activation.

Comparison of A-loop conformation in dimer trajectories

Black – active EGFR

Red – inactive EGFR

Comparison of C-helix conformation in dimer trajectories

Black – active EGFR

Red – inactive EGFR

Comparison of full kinase conformation in dimer trajectories

Black – active EGFR

Red – inactive EGFR

Comparison of C-helix and A-loop conformation in dimer trajectories

RMSD for Yingting’s trajectory

Inactive ActiveFull 4.712377 4.157765alphaC 6.347357 1.874119A-loop 9.82431 4.635911alphaC+A-loop 8.586145 3.768654