A joint experimental and computational study illustrates that the partitioning of positively and negatively charged tryptophan in a phospholipid bilayer is significantly altered by a reversal in the head group dipole arrangement. Experiments were conducted using tryptophan as a fluorescent reporter of its local environment. Based on the experimental design in a recent publication (Anderson, C. M.; Cardenas, A.; Elber, R,; Webb, L. J. J. Phys. Chem. B 2018, 123, 170-179), we were able to determine that the arrangement of the head group dipole altered the degree of partitioning of charged tryptophan in the lipid bilayer. In parallel, atomically detailed simulations were performed for the two membrane systems. The simulation results are in accord with the experimental findings and support a simple molecular partition mechanism of electrostatic interactions with the head groups, glycerol linkers, and interfacial water dipoles.