Force Field Limitations of All-Atom Continuous Constant pH Molecular Dynamics.

All-atom constant pH molecular dynamics simulations offer a powerful tool for understanding pH-mediated and proton-coupled biological processes. As the protonation equilibria of protein sidechains are shifted by electrostatic interactions and desolvation energies, pKa values calculated from the constant pH simulations may be sensitive to the underlying protein force field and water model. Here we investigated the force field dependence of the all-atom particle mesh Ewald (PME) continuous constant pH (PME-CpHMD) simulations of a mini-protein BBL. The replica-exchange titration simulations based on the Amber ff19SB and ff14SB force fields with the respective water models showed significantly overestimated pKa downshifts for a buried histidine (His166) and for two glutamic acids (Glu141 and Glu161) that are involved in salt-bridge interactions. These errors (due to undersolvation of neutral histidines and overstabilization of salt bridges) are consistent with the previously reported \pka's based on the CHARMM c22/CMAP force field, albeit in larger magnitudes. The pKa calculations also demonstrated that ff19SB with OPC water is significantly more accurate than ff14SB with TIP3P water, and the salt-bridge related pKa downshifts can be partially alleviated by the atom-pair specific Lennard-Jones corrections (NBFIX). Together, these data suggest that the accuracies of the protonation equilibria of proteins from constant pH simulations can significantly benefit from improvements of force fields.