Accurately Predicting Protein pK a Values Using Nonequilibrium Alchemy

The stability, solubility, and function of a protein depend on both its net charge and the protonation states of its individual residues. pK(a) is a measure of the tendency for a given residue to (de)protonate at a specific pH. Although pK(a) values can be resolved experimentally, theory and computation provide a compelling alternative. To this end, we assess the applicability of a nonequilibrium (NEQ) alchemical free energy method to the problem of pK(a) prediction. On a data set of 144 residues that span 13 proteins, we report an average unsigned error of 0.77 +/- 0.09, 0.69 +/- 0.09, and 0.52 +/- 0.04 pK for aspartate, glutamate, and lysine, respectively. This is comparable to current state-of-the-art predictors and the accuracy recently reached using free energy perturbation methods (e.g., FEP+). Moreover, we demonstrate that our open-source, pmx-based approach can accurately resolve the pK(a) values of coupled residues and observe a substantial performance disparity associated with the lysine partial charges in Amber14SB/Amber99SB*-ILDN, for which an underused fix already exists.

Automated summary

The stability, solubility, and function of a protein are determined by its net charge and the protonation states of its individual residues. The study demonstrates the applicability of a nonequilibrium alchemical free energy method for accurately predicting pK(a) values, which is comparable to current state-of-the-art predictors. The research also highlights the performance disparity associated with lysine partial charges in certain models and mentions an underused fix for this issue.