Pulling-spring modulation as a method for improving the potential-of-mean-force reconstruction in single-molecule manipulation experiments.

The free-energy calculation is usually limited to close to equilibrium perturbation regimes because faster perturbations introduce a bias into the estimate. The Jarzynski equality offers a solution to this problem by directly connecting the free-energy difference and the external work, regardless how far from equilibrium that work may be. However, a limited sampling coupled to the fast perturbation introduces a slowly converging bias into the Jarzynski free-energy estimate also. In this paper we present two perturbation protocols devised with the intention to overcome the convergence issues of the Jarzynski-based potential of mean force estimation in the single-molecule, constant velocity manipulation experiments. The protocols are designed to improve the convergence issues by increasing the variation of the external work through the modulation of the spring used to pull a molecule. Of the two methods, the one which continuously changes the amplitude of the spring stiffness offers an excellent reconstruction and requires less than one tenth of the samples required by the normal, constant spring pulling to produce the same quality of the reconstruction.