Mechanical Unfolding of Single Polyubiquitin Molecules Reveals Evidence of Dynamic Disorder

[Image: see text] Mechanical unfolding of single polyubiquitin molecules subjected to a constant stretching force showed nonexponentiality in the measured probability density of unfolding (waiting time distribution) and the survival probability of the folded state during the course of the measurements. These observations explored the relevance of disorder present in the system under study with implications for a static disorder approach to rationalize the experimental results. Here, an approach for dynamic disorder is presented based on Zwanzig’s fluctuating bottleneck (FB) model, in which the rate of the reaction is controlled by the passage through the cross-sectional area of the bottleneck. The radius of the latter undergoes stochastic fluctuations that in turn is described in terms of the end-to-end distance fluctuations of the Rouse-like dynamics using a non-Markovian generalized Langevin equation with a memory kernel and Gaussian colored noise. Our results are comprised of analytical expressions for the survival probability and waiting time distribution, which show excellent agreement with the experimental data throughout the range of the applied forces. In addition, by fitting the survival probabilities at different stretching forces, we quantify two system parameters, namely, the average free energy ΔG(av) and the average distance to the transition state Δx(av), both perfectly recovered the experimental estimates. These agreements validate the present model of polymer dynamics, which captures the very essence of dynamic disorder in single-molecule pulling experiments.