Fig. 5. Binding free energy calculation summary for the bromosporine selectivity against 22 bromodomains.

a Calculated vs experimental ΔG values for the HREX FEP¹², FEP without sampling enhancement and non-equilibrium TI approach. In the panels, we also provide a linear regression fit as well as RMSE, AUE, Kendall, Pearson, and Spearman correlations. b Unsigned error (UE) by bromodomain for each of the considered protocols. c Average unsigned error (AUE) with respect to experimental measurement for varying sampling time invested for each of the investigated approaches. The time reflects only the sampling invested in the protein–ligand coupling part of the calculation. The dashed line for the HREX FEP value¹² marks only the accuracy of the estimate using the whole available sampling, i.e., it does not depict dependence on the sampling time. The inset shows the behavior of AUE for FEP and bi-directional non-equilibrium TI when less than 10% of the sampling production time is considered for the ΔG estimation. The uncertainties denote standard errors obtained by bootstrapping.