Figure 4.

Free energy reaction coordinate for two unfolding simulations of GB1. A 1D reaction coordinate is created by calculating the mean distance to reference in a 15-dimensional property space for all structures in an unfolding simulation. The free energy reaction coordinates are calculated by taking the negative log of the counts in a mean distance to the reference histogram. Each 498 K simulation is shown with a black line. The location of unfolding state D is indicated on each free energy reaction coordinate. Top: Run 2 shows three-state unfolding with TS2 located between 0.12 and 0.13 mean distance to reference and TS1 located between 0.23 and 0.25 mean distance to reference. Bottom: Run 4 also shows three-state unfolding with TS2 located between 0.135 and 0.15 mean distance to reference and TS1 located between 0.25 and 0.265 mean distance to reference.