Each complex was solvated in a cubic box of TIP3P water in a box with a buffer of 14 Å from the complex followed by initial minimization for a maximum of 10000 steps using conjugate gradient or until the RMS on the gradient was less than 0.001 kcal/(mol Å). This was followed by 20 ps of NVT and 20 ps of NPT equilibration where all non-water atoms were restrained with a weight of 100 kcal/(mol Å). We finally equilibrated the system for 2 ns in the NPT ensemble without any restraints before our final production run of 10 ns in the NPT ensemble which has been shown to be sufficient in order to obtain reliable results in the subsequent MM/GBSA calculations. The temperature was in all simulations controlled with Langevin dynamics with a collision frequency of γ = 2 ps and the pressure was kept at 1 atm with the weak-coupling algorithm. SHAKE was employed on hydrogen atoms throughout all simulations allowing for a timestep of 2 fs. Structures were saved for postprocessing every 20 ps (500 snapshots) for the analysis of the degree of substitution and every 10 ps (1000 snapshots) for the ensemble simulations for the subsequent calculation of binding free energies with MM/GBSA.