Figure 1.

Concept and workflow of a DFE calculation. (a) One-way trip dissociation of a molecular complex by running metadynamics using an intermolecular distance D as CV. The dotted curve on the right panel is an illustrative trajectory of the centroid of the CV residue of protein B (green ribbon). (b) Primitive FES from a number of replicas of different random seeds, noting that a wall is placed in each run. (c) Average FES stemming from an ensemble of primitive FES, noting the position r₀ of the bound state minimum and the boundary r_b separating the bound state region and the free state region. (d) Boltzmann factor as a function of D, narrowly concentrated around r₀. (e) Convergence analysis: DFE is iteratively calculated as more and more replicas enter into averaging, showing that DFE converges when a sufficient number of replicas are averaged.