To investigate the differences in lipid-protein interactions, we performed simulations of a β 2 -adrenergic receptor (β 2 AR, PDB-ID: 2RH1) embedded in an asymmetric lipid bilayer consisting of 65% POPC and 35% cholesterol in the upper leaflet and 55% POPC, 35% cholesterol, and 10% PIP 2 in the lower leaflet. The initial configuration was obtained from Song et al. (personal communication) and simulated using lincs order = 4, 6, 8 and time steps of ∆t = 10, 20 and 30 fs to assess the impact of improper constraining on the interactions of β 2 AR and cholesterol. Three replicas of 15 µs length were simulated with each combination of lincs order and time step, and the first 5 µs of every trajectory were discarded from analysis as equilibration. All reported quantities were averaged over the three replicas.