We verified that the optimization procedure does not alter the configurations of the interacting beads by running simulations of a single cholesterol molecule in vacuum with both models. Our optimized model excellently reproduces the mean SASA value and its distribution (see Figure ). To further prove the correctness of the optimized model, we computed all pairwise particle distances (Tables and). The optimized model reproduces the mean values of all distances of the original model up to a precision of 0.02 Å, as well as their standard deviation (compare Tables and). Furthermore, we computed the probability density functions of, respectively, the single bond, angle, and dihedral angle of the cholesterol models that do not rely on constraints or virtual sites. The distributions are virtually indistinguishable (Figure ).