Improved Free-Energy Estimates for the Permeation of Bulky Antibiotic Molecules through Porin Channels Using...

The estimation of accurate free energies for antibiotic permeation via the bacterial outer-membrane porins has proven to be challenging. Atomistic simulations of the process suffer from sampling issues that are typical of systems with complex and slow dynamics, even with the application of advanced sampling methods. Ultimately, the objective is to obtain accurate potential of mean force (PMF) for a large set of antibiotics and to predict permeation rates. Therefore, the computational expense becomes an important criterion as well. Simulation studies on the permeation process and similar complex processes have shown that both the sampling scheme employed and the procedure used for the generation of the initial states can critically affect the quality of the estimates obtained and the respective computational overhead. The temperature-accelerated sliced sampling method (TASS) has been shown to partly address the issues with efficient sampling of the important and slow degrees of freedom by enabling simultaneous biasing of a large number of collective variables. In this work, we investigate the effect of the procedure used for the generation of input conformations on the convergence of free-energy estimates obtained from TASS simulations. In particular, we compare the steered molecular dynamics (MD)-based procedure that has been used in previous TASS studies with the Monte Carlo pathway search method, which is used to obtain approximate permeation trajectories with minimum perturbation of the protein channel. We tested different input setups for enrofloxacin permeation through the porins OmpK35 and OmpE35. The best setup shows an improved agreement between independent PMFs in both cases at a much lower computational cost.