Figure 3. Estimation of free energy differences and comparison with experimental measurements.

We divided the global conformational space into two coarse-grained states by defining the separatrix at $S_{path}=0.46$ (0.48 for the triple mutant) in the free energy profile (Figure 2—figure supplement 2) which corresponds to a saddle point of the free energy surface, and then estimated the free energy differences between the two states ($\mathrm{\Delta }G$) from their populations. The time evolution of $\mathrm{\Delta }G$ of L99A (upper time axis) and the triple mutant (lower axis) are shown as black and blue curves, respectively. The experimentally determined values (2.1 kcal mol⁻¹ for L99A and −1.9 kcal mol⁻¹ for the triple mutant) are shown as yellow lines.

DOI: http://dx.doi.org/10.7554/eLife.17505.007