All-atom molecular dynamics simulations of the ion current through OmpF, the major porin in the outer membrane of Escherichia coli, were performed. Starting from the crystal structure, the all-atom modeling allows us to calculate a parameter-free ion conductance in semiquantitative agreement with experiment. Discrepancies between modeling and experiment occur, e.g., at salt concentrations above 1 M KCl or at high temperatures. At lower salt concentrations, the ions have separate pathways along the channel surface. The constriction zone in the channel contains, on one side, a series of positively charges (R42, R82, R132), and on the opposite side, two negatively charged residues (D113, E117). Mutations generated in the constriction zone by removing cationic residues enhance the otherwise small cation selectivity, whereas removing the anionic residues reverses the selectivity. Reduction of the negatively charged residues decreases the conductance by half, whereas cationic residues enhance the conductance. Experiments on mutants confirm the results of the molecular-level simulations.

By reclassifying atoms as QM or MM on-the-fly, adaptive QM/MM dynamics simulations can utilize small QM subsystems whose locations and contents are continuously and automatically updated. Although adaptive QM/MM has been applied in studies of a variety of ions, dynamics simulations of a hydrated proton in bulk water remain a challenge. The difficulty arises from the need to transfer structural features (the covalent and hydrogen bonding networks) via the Grotthuss mechanism instead of the given proton. One must therefore identify an appropriate reference point from which the QM subsystem can be positioned that continuously follows the structural variations as the proton hops. To solve this problem, we propose a proton indicator that serves as the needed reference point. The location of the proton indicator varies smoothly from the hydronium oxygen in the resting (Eigen) state to the shared proton in the transition (Zundel) state. The algorithm is implemented in the framework of a modified permuted adaptive-partitioning QM/MM. As a proof of concept, we simulate an excess proton solvated in bulk water, where the QM subsystem is defined as a sphere of 4.0 Å radius centered at the proton indicator. We find that the use of the proton indicator prevents abrupt changes in the location and contents of the QM subsystem. The new method yields reasonably good agreement in the proton solvation structure and in the proton transfer dynamics with previously reported conventional QM/MM dynamics simulations that employed a much larger QM subsystem (a sphere of 12 Å radius). Also, the results do not change significantly with respect to variations in the time step size (0.1 or 0.5 fs), truncation of the many-body expansion of the potential (from fifth to second order), and absence/presence of thermostat. The proton indicator combined with the modified permuted adaptive-partitioning scheme thus appears to be a useful tool for studying proton transfer in solution.

The temperature-dependent ion conductance of OmpC, a major outer membrane channel of Escherichia coli, is predicted using all-atom molecular dynamics simulations and experimentally verified. To generalize previous results, OmpC is compared to its structural homolog OmpF at different KCl concentrations, pH values, and a broad temperature range. At low salt concentrations and up to room temperature, the molecular modeling predicts the experimental conductance accurately. At high salt concentrations above 1 M KCl and above room temperature, the simulations underestimate the conductance. Moreover, the temperature dependence of the channel conductance is different from that of the bulk, both in experiment and simulation, indicating a strong contribution of surface effects to the ion conductance. With respect to OmpC, subconductance levels can be observed in experiments only. Subconductance and gating levels can be clearly distinguished by their differences in conductance values and temperature-dependent behavior. With increasing temperature, the probability of a subconductance state to occur, increases, while the dwell time is decreased. The open probability, frequency, and dwell time of such states is largely pH- and KCl concentration-independent, while their amplitudes show a lower increase with increasing salt concentration than gating amplitudes. Voltage dependence of subconductance has been found to be negligible within the uncertainty of the measurements.

The adaptive-partitioning (AP) schemes for combined quantum-mechanical/molecular-mechanical (QM/MM) calculations allow on-the-fly reclassifications of atoms and molecules as QM or MM in dynamics simulations. The permuted-AP (PAP) scheme (J. Phys. Chem. B 2007, 111, 2231) introduces a thin layer of buffer zone between the QM subsystem (also called active zone) and the MM subsystem (also known as the environmental zone) to provide a continuous and smooth transition and expresses the potential energy in a many-body expansion manner. The PAP scheme has been successfully applied to study small molecules solvated in bulk solvent. Here, we propose two modifications to the original PAP scheme to treat solvent molecules entering and leaving protein binding sites. First, the center of the active zone is placed at a pseudoatom in the binding site, whose position is not affected by the movements of ligand or residues in the binding site. Second, the extra forces due to the smoothing functions are deleted. The modified PAP scheme no longer describes a Hamiltonian system, but it satisfies the conservation of momentum. As a proof-of-concept experiment, the modified PAP scheme is applied to the simulations under the canonical ensemble for two binding sites of the Escherichia coli CLC chloride ion transport protein, in particular, the intracellular binding site Sint discovered by crystallography and one putative additional binding site Sadd suggested by molecular modeling. The exchange of water molecules between the binding sites and bulk solvent is monitored. For comparison, simulations are also carried out using the same model system and setup with only one exception: the extra forces due to the smoothing functions are retained. The simulations are benchmarked against conventional QM/MM simulations with large QM subsystems. The results demonstrate that the active zone centered at the pseudo atom is a reasonable and convenient representation of the binding site. Moreover, the transient extra forces are non-negligible and cause the QM water molecules to move out of the active zone. The modified PAP scheme, where the extra forces are excluded, avoids the artifact, providing a realistic description of the exchange of water molecules between the protein binding sites and bulk solvent.

The semiempirical MNDO-based AM1 and PM3 methods and the orthogonalization-corrected OM1, OM2, and OM3 models were reparameterized to improve their description of bulk water and of proton transfer in water. Reference data included the gas-phase geometries and energies of the water molecule, small water clusters, the hydronium ion, and small hydronium ion-water clusters, as well as the gas-phase potential energy surface for proton transfer between the two water molecules in a Zundel ion, all calculated at the MP2/aug-cc-pVTZ level of theory. Combined QM/MM molecular dynamics simulations were carried out for bulk water and for a proton solvated in water using large cluster models. Both the authentic and reparameterized semiempirical models were employed in the simulations. The reparameterization led to significantly better results in all cases. The new set of OM3 parameters gave the best overall results for the structural and dynamic properties of water and the hydrated proton, with a small but finite barrier of 0.1-0.2 kcal/mol in the potential of mean force for proton transfer, in agreement with ab initio path-integral molecular dynamics simulations. The reparameterized OM3 model is expected to be useful for efficient modeling of proton transfer in aqueous solution.