Our choice of running 25 simulations of 4 nanoseconds each is in accordance with previous findings by our group showing that 25 replicas of 4 ns are sufficient to obtain converged values of ∆G ESM ACS . These trajectories were post-processed in MMPBSA.py to produce 25 binding free energy estimates, one for each replica within the ensemble. The reported ∆G ESM ACS is the mean of the sampling distribution of means for this sample of 25 free energy estimated obtained using bootstrapping. The associated error bars are the corresponding standard errors.