Pressure-induced phase transitions of lipid bilayers observed by fluorescent probes Prodan and Laurdan Original Research Article

The fluorescence spectra of 6-propionyl-2-(dimethylamino)naphthalene (Prodan) and 6-dodecanoyl-2-(dimethylamino)naphthalene (Laurdan) in bilayer membranes of 1,2-distearoylphosphatidylcholine (DSPC) were observed as a function of pressure at constant temperature. The emission spectra of Prodan and Laurdan varied with the pressure-induced states of bilayer membranes. The maximum emission wavelength (λmax) of Prodan characteristic of the liquid crystalline (Lα), lamellar gel (Lβ′) and pressure-induced interdigitated gel (LβI) phases of the DSPC bilayer was 480, 440 and 500 nm, respectively. On the other hand, the λmax of Laurdan characteristic of the Lα and Lβ′ phases was 480 and 440 nm in a similar manner as Prodan probe. However, no change in the λmax was observed in spite of the occurrence of the interdigitation of bilayer. Since the λmax reflects the solvent property around the probe molecules, we could speculate about the location of fluorescent probe in the bilayer membranes. In the Lα phase the same chromophore group of Prodan and Laurdan probes distributes around phosphate group of lipid (i.e., polar region). The transformation of bilayer into the Lβ′ phase causes the Prodan and Laurdan molecules to move into the glycerol backbone (i.e., less polar) region. In the ripple gel (Pβ′) phase, the emission spectrum of Prodan shows a broad peak at about 480 nm and a shoulder around 440 nm, which means that the Prodan molecules are widespread over the wide range from the glycerol backbone to the hydrophilic part of bilayer. The Pβ′ / LβI phase transition causes the Prodan molecule to squeeze out from the glycerol backbone region and to move the hydrophilic region near the bilayer surface. Contrarily, the Laurdan molecule was not squeezed out from the glycerol backbone region because the long acyl chain of Laurdan serves as an anchor in the hydrophobic core of bilayer. The ratio of fluorescence intensity of Prodan at 480 nm to that at 440 nm, F480 / F440, is available to observation of bilayer phase transitions. The plot of F480 / F440 versus pressure seems to be useful for the recognition of bilayer phase transition, especially the bilayer interdigitation.