Beyond microscopic reversibility: Are observable non-equilibrium processes precisely reversible?

Although the principle of microscopic reversibility has been studied for many decades, there remain ambiguities in its application to non-equilibrium processes of importance to chemistry, physics and biology. Examples include whether protein unfolding should follow the same pathways and in the same proportions as folding, and whether unbinding should likewise mirror binding. Using continuum-space calculations which extend previous kinetic analyses, we demonstrate formally that the precise symmetry of forward and reverse processes is expected only under certain special conditions. Approximate symmetry will be exhibited under a separate set of conditions. Exact, approximate, and broken symmetry scenarios are verified in several ways: using numerical calculations on toy and molecular systems; using exact calculations on kinetic models of induced fit in protein-ligand binding; and based on reported experimental results. The analysis highlights intrinsic challenges and ambiguities in the design and analysis of both experiments and simulations.