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  • Title: Exploration of physical principles underlying lipid regular distribution: effects of pressure, temperature, and radius of curvature on E/M dips in pyrene-labeled PC/DMPC binary mixtures.
    Author: Chong PL, Tang D, Sugar IP.
    Journal: Biophys J; 1994 Jun; 66(6):2029-38. PubMed ID: 8075336.
    Abstract:
    In a previous study, we observed a series of dips in the plot of E/M (the ratio of excimer to monomer fluorescence intensity) versus the mole fraction of 1-palmitoyl-2-(10-pyrenyl)decanoyl-sn-glycerol-3-phosphatidylcholine (Pyr-PC) in Pyr-PC/DMPC binary mixtures at 30 degrees C. In the present study, we have characterized the physical nature of E/M dips in Pyr-PC/DMPC binary mixtures by varying pressure, temperature, and vesicle diameter. The E/M dips at 66.7 and at 71.4 mol% PyrPC in DMPC multilamellar vesicles remain discernible at 30-43 degrees C. At higher temperatures (e.g., 53 degrees C), the depth of the dip abruptly becomes smaller. This result agrees with the idea that E/M dips appear as a result of regular distribution of pyrene-labeled acyl chains into hexagonal super-lattices at critical mole fractions. Regular distribution is a self-ordering phenomenon. Usually, in self-ordered systems, the number of structural defects increases with increasing temperature, and thermal fluctuations eventually result in an order-to-disorder transition. The effect of vesicle diameter on the E/M dip at 66.7 mol% Pyr-PC in DMPC has been studied at 37.5 degrees C by using unilamellar vesicles of varying sizes. The E/M dip is observable in large unilamellar vesicles; however, the depth of the E/M dip decreases when the vesicle diameter is reduced. When the vesicle diameter is reduced to about 64 nm, the dip becomes shallow and split. This result suggests that the curvature-induced increase in the separation of lipids in the outer monolayer decreases the tendency of regular distribution for pyrene-labeled acyl chains. Regular distribution is believed to arise from the long-range repulsive interaction between Pyr-PC molecules due to the elastic deformation of the lipid matrix around the bulky pyrene moiety. When the radius of curvature becomes small, outer monolayer lipids are more separated. Therefore, pyrene-containing acyl chains fit better into the membrane matrix, which alleviates the deformation of the lattice and diminishes the long-range repulsive interactions between pyrene-containing acyl chains. Furthermore, we have shown a striking difference in the pressure dependence of E/M at critical Pyr-PC mole fractions and at noncritical mole fractions. In the pressure range between 0.001 and 0.7 kbar at 30 degrees C, E/M decreases steadily with increasing pressure at noncritical mole fractions; in contrast, E/M changes little with pressure at critical mole fractions (e.g., 33.3 and 50.0 mol% Pyr-PC). The pressure data suggest that membrane free volume in the liquid crystalline state of the bilayer is less abundant at critical Pyr-PC mole fractions than at noncritical mole fractions.
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