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Single-molecule observations of biomolecular dynamics and folding are commonly rationalized using the model of diffusive dynamics on a free-energy landscape, which is inferred via the Boltzmann inversion of the equilibrium distribution of the experimental observable. Can the same model be applied to high-resolution single-molecule trajectories of molecular machines that lack thermal equilibrium so that the Boltzmann inversion method is inapplicable? In this Letter, we discuss two approaches to reconstructing the underlying free-energy landscape in such nonequilibrium systems and explore the performance of this model in application to trajectories with complex underlying dynamics.
Berezhkovskii et al. (Tue,) studied this question.
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