It is, however, important to understand the limitations of the ∆G values obtained this way. The calculated values should not be interpreted as reporting on the actual free energy differences between the apo conformers, but only on a component of ∆G corresponding to the change in the degrees of freedom relevant for ligand binding. It is likely that the binding site rearrangements are experienced by the ligands and have a strong effect on the ∆G calculated based on this approach. At the same time, substantial conformational rearrangements further from the binding site may not have a contribution to ∆G if they do not affect the ligand binding affinity. Therefore, the conclusions about the most likely apo state identified with this approach should be limited to the interpretations of the binding affinities for a specific set of ligands.