The aliphatic chain dynamics of our atomistic cholesterol reference could not be fully captured when using a single angle potential along beads R5-C1-C2, as was the case with Martini 2, and instead required a dihedral potential in place. This is likely due to the differences in interaction radii between the different Martini 3 bead sizes that allow for closer interactions and increased configurational freedom compared to Martini 2 . The R6-R4-R3-C2 improper dihedral and R4-R3-C2 angle potentials were added in place with enough flexibility to account for the dynamics of cholesterol both in a lipid membrane and in solution (Supplementary Figure ). The potentials were selected so as to minimize the probability of issues from numerical instabilities arising from angles associated with dihedrals reaching 0 or 180 degrees .