Additionally, SMD was used to enforce the effects of the E/Zphotoisomerization of CCBu. This was done by using the equilibrated systems involving 5, 10, 15, 20, 30, 40 and 50 CCBu (corresponding to a CCBu/lipids ratio of 0.25, 1.25, 2.5, 3.75, 5, 7.5, 10, 12.5%) in E-conformation within the lipid bilayer and applying a 5 kcal/mol harmonic potential to force the dihedral angle around the isomerizable carbon-carbon double bond to change from 179° (E-isomer) to 7.9° (Z-isomer). The harmonic potential was applied for 100,000 steps using a time step of 0.5 fs to avoid numerical instabilities. After the dihedral angle change, the force fields of CCBu was switched to the one describing its Z-conformation and unconstrained equilibrium MD simulation performed for 1130 ns.