Quantitative contributions of cholesterol and the individual classes of phospholipids and their degree of fatty acyl (un)saturation to membrane fluidity measured by fluorescence polarization.
Steady-state fluorescence polarization (P) measurements, using the probe 1,6-diphenyl-1,3,5-hexatriene, in a large variety of well-defined liposomes at 25 degrees C allowed a quantitative description of the contributions of cholesterol, sphingomyelin, and (un)saturation of fatty acyl groups in the various phospholipids to the structural order (or the mutual affinity) of membrane lipids. The P values for liposomes prepared from lipid extracts of natural (purified) membranes of various origins could be more or less predicted (calculated) from the relative contributions of the individual lipid components. In all cases, the polarization varied with the cholesterol/phospholipid molar ratio (C/PL) according to the equation P = Pplat -(Pplat -Pzero) exp(-alpha C/PL), in which Pzero refers to the polarization without cholesterol and Pplat is a maximal plateau value, reached at a high C/PL (greater than 1). The "cholesterol-ordering coefficient" alpha of the phospholipids was found to increase with the fraction of sphingomyelin or dipalmitoylphosphatidylcholine molecules, indicating that the susceptibility of phospholipids to be ordered by cholesterol is increased by these compounds. Pzero increases curvilinearly with the fraction of either of these molecules. Pplat increases linearly with the fraction of saturated acyl chains for most phospholipids. Highly unsaturated fatty acyl chains (e.g., 20:4 and 22:6) strongly depress Pplat but not Pzero. The results suggest that such phospholipids are unlikely to associate with cholesterol and may thus create extremely fluid membrane domains. The disproportionation of cholesterol in the cell can be understood by the differing composition of the phospholipids in plasma membranes and endomembranes and their ordering susceptibility (affinity) toward cholesterol.
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D006801:Humans; D007700:Kinetics; D007942:Leukemia, Experimental / Q000378:metabolism; D008081:Liposomes; D008099:Liver / Q000378:metabolism; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D010713:Phosphatidylcholines; D010743:Phospholipids / Q000378:metabolism; D051381:Rats; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D008081:Liposomes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D002784:Cholesterol