The interaction of cholesterol and cholest-4-en-3-one with dipalmitoylphosphatidylcholine. Comparison based on the use of three fluorophores

This study compares cholesterol-phospholipid and cholest-4-en-3-one-phospholipid interactions by their effect on thermotropic behavior of dipalmitoylphosphatidylcholine bilayers. This was approached by determining the temperature-dependent steady-state fluorescence anisotropy of three fluorophores; diphenylhexatriene (DPH), hydroxycoumarin (HC) and trans-parinaric acid (TPA). The fluorophores monitor different lateral and vertical locations of the lipid bilayers; DPH and HC average laterally the properties of the hydrophobic and headgroup regions of the bilayer, respectively, while TPA distribution is determined by the lateral organization of the bilayer. The data show that the two steroids have similar qualitative but different quantitative effects. Both diminish the pretransition and behave as 'averagers', broadening the main gel to liquid crystalline phase transition through ordering of the acyl chains in the liquid crystalline state and disordering of them in the gel state. However, the mechanisms by which the two molecules operate are different. Cholesterol is more effective particularly on the hydrophobic region of the bilayer, and its effect is not linear with its mole fraction. A sharp increase of the steady-state fluorescence anisotropy occurs around 20 mol% cholesterol. The effect of cholestenone is proportional to its mole fraction. The difference between the effects of the two steroids is explained by the dissimilarity in their lateral distribution. Cholesterol forms cholesterol-rich domains. The size of the boundary regions which surround the cholesterol-rich domains changes drastically at about 20 mol% cholesterol. Cholestenone, on the other hand, is randomly distributed in the bilayer plane and therefore it does not cause the formation of such defined boundary regions. This study as well as reports by others suggests that the important structural differences between the two steroids are the molecular packing parameter and the presence of small polar group at the 3β position of the steroid.