Erratum: Ratchet universality in the bidirectional escape from a symmetric potential well [Phys. Rev. E 103, 022203 (2021)].

This corrects the article DOI: 10.1103/PhysRevE.103.022203.

Ratchet universality in the bidirectional escape from a symmetric potential well.

The present work discusses symmetry-breaking-induced bidirectional escape from a symmetric metastable potential well by the application of zero-average periodic forces in the presence of dissipation. We characterized the interplay between heteroclinic instabilities leading to chaotic escape and breaking of a generalized parity symmetry leading to directed ratchet escape to an attractor either at ∞ or at -∞. Optimal enhancement of directed ratchet escape is found to occur when the wave form of the zero-average periodic force acting on the damped driven oscillator matches as closely as possible to a universal wave form, as predicted by the theory of ratchet universality. Specifically, the optimal approximation to the universal force triggers the almost complete destruction of the nonescaping basin for driving amplitudes which are systematically lower than those corresponding to a symmetric periodic force having the same period. We expect that this work could be potentially useful in the control of elementary dynamic processes characterized by multidirectional escape from a potential well, such as forced chaotic scattering and laser-induced dissociation of molecular systems, among others.

Interrelationships between tyrocidine and gramicidin A' in their interaction with phospholipids in model membranes.

(1) The interaction of tyrocidine with different lipids is studied in model membranes and the results are compared to the gramicinid-lipid interaction. (2) The tyrocidine-dielaidoylphosphatidylethanolamine interaction gives rise to a population of phospholipids with a lower gel to liquid-crystalline transition temperature and to an abolition of the bilayer to HII phase transition, resulting in a macroscopic organization with dynamic and structural properties different from those of the pure lipid. (3) Tyrocidine has a strong fluidizing effect on the acyl chains of phosphatidylcholines, manifested by a decrease in enthalpy of the main thermotropic transition. (4) No evidence of a gramicidin A'-like lipid-structure modulating activity was found. However, tyrocidine inhibits the formation by gramicidin of an HII phase in dioleoylphosphatidylcholine model membranes. Instead, a cubic type of lipid organization is observed. (5) Tyrocidine greatly perturbs the barrier properties of dioleoylphosphatidylcholine model membrane. (6) Gramicidin A' reverses the effect of tyrocidine on membrane permeability by forming a complex in the model membrane with an apparent 1:1 stoichiometry. (7) The results suggest that both peptide antibiotics, which are produced by Bacillus brevis ATC 8185 prior to sporulation, show antagonism in their effect on membrane structure similar to their effect on superhelical DNA (Bogh, A. and Ristow, H. (1986) Eur. J. Biochem. 160, 587-591. The possible underlying basic mechanism is indicated.

The interaction of abietic acid with phospholipid membranes.

Abietic acid is a major component of the oleoresin synthesized by many conifers and constitutes a major class of environmental toxic compounds with potential health hazard to animal, including human, and plant life. Being an amphipathic molecule, the study of the influence of abietic acid on the structure of membranes would be important to get insight into the mechanism of toxic action of the molecule. The interaction of abietic acid with model membranes of dipalmitoylphosphatidylcholine (DPPC) and dielaidoylphosphatidylethanolamine (DEPE) has been studied by differential scanning calorimetry and 31P-nuclear magnetic resonance spectroscopy. It has been found that abietic acid greatly affects the phase transition of DPPC, shifting the transition temperature to lower values, giving rise to the appearance of two peaks in the thermogram and to the presence of fluid immiscible phases. In a similar way, the phase transition of DEPE, in the presence of abietic acid, was shifted to lower temperatures, and two peaks appeared in the thermograms. The temperature of the lamellar to hexagonal H(II) phase transition was also decreased by the presence of abietic acid, but phase immiscibilities were not detected. The possible implications of these effects on the action of abietic acid on biological membranes are discussed.

The influence of vitamin K1 on the structure and phase behaviour of model membrane systems.

Vitamin K1 is a component of the Photosystem I of plants which constitutes the major dietary form of vitamin K. The major function of this vitamin is to be cofactor of the microsomal gamma-glutamylcarboxylase. Recently, novel roles for this vitamin in the membrane have been postulated. To get insight into the influence of vitamin K1 on the phospholipid component of the membrane, we have studied the interaction between vitamin K1 and model membranes composed of dimyristoylphosphatidylcholine (DMPC) and dielaidoylphosphatidylethanolamine (DEPE). We utilized high-sensitivity differential scanning calorimetry and small-angle X-ray diffraction techniques. Vitamin K1 affected the thermotropic properties of the phospholipids, broadened and shifted the transitions to lower temperatures, and produced the appearance of several peaks in the thermograms. The presence of the vitamin gave rise to the formation of vitamin-rich domains which were immiscible with the bulk phospholipid in both the gel and the liquid-crystalline phases. Vitamin K1 was unable to alter the lamellar organization of DMPC, but we found that it produced an increase in the interlamellar repeat spacing of DMPC at 10 degrees C. Interestingly, vitamin K1 promoted the formation of inverted hexagonal HII structures in the DEPE system. We discuss the possible implications that these vitamin K1-phospholipid interactions might have with respect to the biological function of the vitamin.

A fluorescence and microcalorimetric study of the interaction between lasalocid A and phospholipid vesicles.

The binding of lasalocid A to dipalmitoylphosphatidylcholine (DPPC) vesicles was studied following changes in the intrinsic fluorescence of this ionophore. The binding calculations indicated a dissociation constant of 6.98 +/- 1.5 muM at 48 degrees C, i.e., above the transition temperature (Tc) of the pure phospholipid, with a number of binding sites of 1 per 22 +/- 2.5 molecules of phospholipid, while at 23 degrees C, i.e., below the Tc of the pure phospholipid, the dissociation constant was 9.15 +/- 0.24 muM and the number of binding sites was 1 per each 29 +/- 1.6 molecules of DPPC. Changes in the temperature induced changes in fluorescence intensity of lasalocid A mainly upon phase changes, indicating a progressive decrease in the transition temperature accompanied by a broadening of the transition as lasalocid A concentration was increased. Fluorescence quenching experiments with N-methylnicotinamide showed a certain accessibility of the fluorophoric group of the ionophore to the aqueous quencher. Differential scanning calorimetry showed that increasing concentrations of lasalocid A drastically modified the thermotropic profile. At concentrations higher than 5 mol%, a second peak appeared, possibly due to a lateral phase segregation of lasalocid A trapping some phospholipid molecules. The results are interpreted in terms of limited solubility of lasalocid A in the phospholipid vesicles, this solubility being higher in fluid than in rigid phospholipid. Lateral segregation seems to occur with formation of more than one phase. At least the salicylic acid residue of the ionophore appears to be located near the polar head group of the phospholipid.

A differential scanning calorimetry study of the interaction of alpha-tocopherol with mixtures of phospholipids.

When alpha-tocopherol was included in multibilayer vesicles of dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine and distearoylphosphatidylcholine it induced a broadening of the main transition and a displacement of this transition to lower temperatures, as seen by differential scanning calorimetry. This effect was quantitatively more important in the samples of distearoylphosphatidylcholine than in those of the other phosphatidylcholines. Alpha-Tocopherol when present in equimolar mixtures of dimyristoylphosphatidylcholine and diastearoylphosphatidylcholine, which show monotectic behaviour, preferentially partitions in the most fluid phase. The effect of alpha-tocopherol on the phase transition of dilauroylphosphatidylethanolamine and dipalmitoylphosphatidylethanolamine is qualitatively different of that observed on phosphatidylcholines, and several peaks are observed in the calorimetric profile, probably indicating the formation of separated phases with different contents in alpha-tocopherol. The effect was more apparent in dipalmitoylphosphatidylethanolamine than in dilauroylphosphatidylethanolamine. The inclusion of alpha-tocopherol in equimolar mixtures of dilauroylphosphatidylethanolamine and dipalmitoylphosphatidylcholine, which show cocrystallization, only produced a broadening of the phase transition and a shift to lower temperatures. However, in the case of equimolar mixtures of dipalmitoylphosphatidylcholine which also show cocrystallization, the effect was to cause lateral phase separation with the formation of different mixtures of phospholipids and alpha-tocopherol. Alpha-Tocopherol was also included in equimolar mixtures of phosphatidylethanolamine and phosphatidylcholine showing monotectic behaviour, and in this case alpha-tocopherol preferentially partitioned in the most fluid phase, independently of whether this was composed mainly of phosphatidylcholine or of phosphatidylethanolamine.

Importance of the tryptophans of gramicidin for its lipid structure modulating activity in lysophosphatidylcholine and phosphatidylethanolamine model membranes. A comparative study employing gramicidin analogs and a synthetic alpha-helical hydrophobic polypeptide.

The importance of the tryptophan residues of gramicidin for the lipid structure modulating activity of this pentadecapeptide was investigated by studying the interaction of gramicidin analogs A, B, C (which have a tryptophan, phenylalanine and tyrosine in position 11, respectively) and tryptophan-N-formylated gramicidin (in which the four tryptophan residues have been formylated) with several phospholipid systems. In addition an alpha-helical model pentadecapeptide (P15) was studied to further test the specificity of the gramicidin-lipid interaction. DSC experiments showed that all the gramicidin analogs produced a significant decrease in the gel to liquid-crystalline transition enthalpy of dipalmitoylphosphatidylcholine. The P15 peptide was much less effective in this respect. In dielaidoylphosphatidylethanolamine the gel----liquid-crystalline transition enthalpy was much less affected by the incorporation of these molecules. In this lipid system tryptophan-N-formylated gramicidin was found to be the most ineffective. 31P-NMR and small angle X-ray diffraction experiments showed that the ability of the peptides to induce bilayer structures in palmitoyllysophosphatidylcholine and HII phase promotion in dielaidoylphosphatidylethanolamine systems follows the order: gramicidin A' (natural mixture) approximately equal to gramicidin A greater than gramicidin B approximately equal to gramicidin C greater than tryptophan-N-formylated gramicidin greater than P15. These results support the hypothesis that the shape of gramicidin and its aggregational behaviour, in which the tryptophan residues play an essential role, are major determinants in the unique lipid structure modulating activity of gramicidin.

Capsaicin affects the structure and phase organization of phospholipid membranes.

Capsaicin is a natural compound with pharmacological and toxicological effects, which given its hydrophobicity, can influence the structure of membranes. The interaction of capsaicin with model membranes of dipalmitoylphosphatidylcholine and dielaidoylphosphatidylethanolamine has been studied by using differential scanning calorimetry, fluorescent probe spectroscopy and 31P-nuclear magnetic resonance. Capsaicin remarkably affects the phase transition of dipalmitoylphosphatidylcholine, shifting the transition temperature to lower values, and giving rise, at relatively high capsaicin concentrations, to the appearance of two peaks in the thermogram. These peaks may correspond to separated phases as indicated by the partial phase diagram. Whereas capsaicin did not affect the fluorescence polarization of the probes diphenylhexatriene and trimethylammonium-diphenylhexatriene, it clearly affected that of the probe 2-anthroyloxystearic acid, indicating that the perturbation produced by capsaicin on the membrane would be mainly at the position where this fluorophore is located. On the other hand, capsaicin, at relatively low concentrations, gives rise to immiscible phases in the presence of dielaidoylphosphatidylethanolamine and decrease the temperature of the lamellar to hexagonal HII phase transition. At concentrations of capsaicin higher than 0.3 mol fraction, isotropic phases were detected. The possible implications of the effects of capsaicin on biological membranes are discussed.

Interaction of retinol and retinoic acid with phospholipid membranes. A differential scanning calorimetry study.

The influence of retinol and retinoic acid, two retinoids of major interest, on the main gel to liquid-crystalline phase transition of different phospholipid membranes has been studied by means of differential scanning calorimetry. Both compounds exerted perturbing effects on the phase transition of membranes composed of dipalmitoylphosphatidylcholine or dipalmitoylphosphatidylethanolamine. At concentrations up to 42.5 mol% of retinoid in the membrane, the delta H was not much affected with respect to the pure phospholipid, indicating a rather slight interaction. As the concentration of retinol was increased the Tc transition temperature decreased. A fluid-phase immiscibility was observed for the system DPPC/retinol at concentrations between 0 and 33 mol%. Almost ideal phase diagrams were obtained for the mixture DPPE/retinol. At concentrations of 33 mol% and higher retinol was able to induce phase separations in DPPC membranes, but not in DPPE. The effect of retinoic acid was much weaker, the Tc and delta H remaining almost unaltered and equal to that of the pure phospholipid up to concentrations of 30 mol%, at neutral pH. Retinoic acid exerted a pH-dependent effect. As the pH decreased, and therefore increased the extent of protonation of retinoic acid, the pertubation of the membrane induced by this compound was less. A strong effect, both on Tc and delta H, was observed at pH 10, where the retinoic acid moiety will be mainly unprotonated and the negative charge will generate repulsive forces thus destabilizing the membrane. The mixture DPPC/retinoic acid presents a region of fluid-phase immiscibility. At low pH, when the retinoic acid moiety was fully protonated, this fluid-immiscibility region extended from 0 to 36 mol% of retinoic acid, but its size decreased with increasing pH, and at pH 10 it was only found from 0 to 3 mol%. These results are discussed in terms of the possible retinoid/phospholipid interactions and the disposition of the retinoid moiety in the bilayer.

Influence of retinoids on phosphatidylethanolamine lipid polymorphism.

The interaction of all-trans-retinoic acid and all-trans-retinol with dielaidoylphosphatidylethanolamine has been studied by differential scanning calorimetry and 31P-NMR spectroscopy. Increasing concentrations of all-trans-retinoic acid up to a mol fraction of 0.09 were found to induce shifts to lower temperatures of both the L beta to L alpha and L alpha to hexagonal-HII phase transitions, with a slight decrease in the enthalpy change of the transitions. At higher concentrations no further effects on the transitions were observed, and this is interpreted as indicative of a limited miscibility of retinoic acid with the phospholipid. 31P-NMR spectroscopy confirmed that the L alpha to hexagonal-HII phase transition was shifted to lower temperatures in the presence of retinoic acid. On the other hand increasing concentrations of all-trans-retinol up to a mol fraction of 0.166, induced a progressive shift of the L beta to L alpha and the L alpha to hexagonal-HII phase transitions to lower temperatures. At higher concentrations the main gel to liquid-crystalline phase transition was further displaced to lower temperatures and the lamellar to hexagonal-HII phase transition was not observed in the thermograms. 31P-NMR spectroscopy indicated that retinol was able of inducing the phospholipid to adopt the hexagonal-HII phase at temperatures even below the main gel to liquid-crystalline phase transition temperature of the pure phospholipid.

The interaction of alpha-tocopherol with phosphatidylserine vesicles and calcium.

The interaction of alpha-tocopherol with dimyristoylphosphatidylserine (DMPS) has been studied in the presence and in the absence of Ca2+ by using differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR) and 45Ca2+-binding. In the absence of Ca2+, DSC showed that alpha-tocopherol decreases the temperature of the lamellar gel to lamellar liquid crystalline phase transition as well as it decreases delta H of this transition. Two different peaks were detected at 10 mol% of alpha-tocopherol and probably one of the peaks correspond to pure or nearly pure DMPS and the other to DMPS incorporating most of the alpha-tocopherol. The phase transition was totally abolished at 30 mol% of alpha-tocopherol. In the presence of Ca2+ this L(beta) to L(alpha) phase transition of DMPS was even more perturbed by alpha-tocopherol, so that it was totally abolished by only 7 mol% of alpha-tocopherol, at Ca2+ concentrations which were clearly non-saturating, like those giving DMPS/Ca2+ molar ratio of 4:1 and 10:1. Furthermore, the transition of the DMPS/Ca2+ complex observed at 91.6 degrees C was perturbed by the presence of alpha-tocopherol, indicating a change in the structure of the crystalline complex. The FT-IR analysis of the effect of alpha-tocopherol on DPMS phase transition confirmed the decrease in the phase transition temperature of the phospholipid, and also that alpha-tocopherol increases the number of gauche isomers in the gel state but has no effect in the liquid crystalline state. The binding of 45Ca2+ was also affected by the presence of alpha-tocopherol, so that the number of binding sites was decreased, and this may be interesting for situations in which phosphatidylserine and Ca2+ are simultaneously implicated in biological functions, such as membrane fusion and enzyme activation.

Influence of vitamin E on phosphatidylethanolamine lipid polymorphism.

The effect of vitamin E, in its major form alpha-tocopherol and its synthetic analog alpha-tocopheryl acetate, on phosphatidylethanolamine lipid polymorphism has been studied by mean of differential scanning calorimetry and 31P-nuclear magnetic resonance techniques. From the interaction of these tocopherols with dielaidoylphosphatidylethanolamine it is concluded that both molecules promote the formation of the hexagonal HII phase at temperatures lower than those of the pure phospholipid. When the tocopherols were incorporated in the saturated dimiristoylphosphatidylethanolamine, which has been shown not to undergo bilayer to hexagonal HII phase transition, up to 90 degrees C, they induce the phospholipid to partially organize in hexagonal HII phase. From our experiments it is shown that alpha-tocopherol is more effective than its analog in promoting HII phase in these systems. It is also shown that, while alpha-tocopheryl acetate does not significantly perturb the gel to liquid-crystalline phase transition of dimirystoylphosphatidylethanolamine, alpha-tocopherol does so and more than one peak appears in the calorimetric profile, indicating that lateral phase separations are taking place.

Effects of (+)-totarol, a diterpenoid antibacterial agent, on phospholipid model membranes.

(+)-Totarol, a highly hydrophobic diterpenoid isolated from Podocarpus spp., is inhibitory towards the growth of diverse bacterial species. (+)-Totarol decreased the onset temperature of the gel to liquid-crystalline phase transition of DMPC and DMPG membranes and was immiscible with these lipids in the fluid phase at concentrations greater than 5 mol%. Different (+)-totarol/phospholipid mixtures having different stoichiometries appear to coexist with the pure phospholipid in the fluid phase. At concentrations greater than 15 mol% (+)-totarol completely suppressed the gel to liquid-crystalline phase transition in both DMPC and DMPG vesicles. Incorporation of increasing amounts of (+)-totarol into DEPE vesicles induced the appearance of the H(II) hexagonal phase at low temperatures in accordance with NMR data. At (+)-totarol concentrations between 5 and 35 mol% complex thermograms were observed, with new immiscible phases appearing at temperatures below the main transition of DEPE. Steady-state fluorescence anisotropy measurements showed that (+)-totarol decreased and increased the structural order of the phospholipid bilayer below and above the main gel to liquid-crystalline phase transition of DMPC respectively. The changes that (+)-totarol promotes in the physical properties of model membranes, compromising the functional integrity of the cell membrane, could explain its antibacterial effects.

Organotin compounds alter the physical organization of phosphatidylcholine membranes.

Organotin compounds have a broad range of biological activities and are ubiquitous contaminants in the environment. Their toxicity mainly lies in their action on the membrane. In this contribution we study the interaction of tributyltin and triphenyltin with model membranes composed of phosphatidylcholines of different acyl chain lengths using differential scanning calorimetry, (31)P-nuclear magnetic resonance, X-ray diffraction and infrared spectroscopy. Organotin compounds broaden the main gel to liquid-crystalline phase transition, shift the transition temperature to lower values and induce the appearance of a new peak below the main transition peak. These effects are more pronounced in the case of tributyltin and are quantitatively larger as the phosphatidylcholine acyl chain length decreases. Both tributyltin and triphenyltin increase the enthalpy change of the transition in all the phosphatidylcholine systems studied except in dilauroylphosphatidylcholine. Organotin compounds do not affect the macroscopic bilayer organization of the phospholipid but do affect the degree of hydration of its carbonyl moiety. The above evidence supports the idea that organotin compounds are located in the upper part of the phospholipid palisade near the lipid/water interface.

Interaction between alpha-tocopherol and heteroacid phosphatidylcholines with different amounts of unsaturation.

Differential scanning calorimetry was used to study the influence of a alpha-tocopherol on the thermotropic properties of model membranes composed by a series of heteroacid phosphatidylcholines with different amount of unsaturation in the sn-2 chain. The effect of alpha-tocopherol on 1,2-distearoylglycerophosphocholine (18:0,18:0), 1-stearoyl-2-oleoylgylcerophosphocholine (18:0,18:1), 1-stearoyl-2-linoleoylglycerophosphocholine (18:0,18:2), 1-stearoyl-2-linolenoylglycerophosphocholine (18:0,18:3), and 1-stearoyl-2-arachidonoylglycerophosphocholine (18:0,20:4) was determined. The addition of alpha-tocopherol perturbed the thermotropic gel to liquid-crystalline phase transition of these phospholipids. alpha-Tocopherol broadened the endotherm, lowered the transition temperature and decreased the associated enthalpy change. Partial phase diagrams showed the presence of fluid immiscibilities giving rise to lateral phase separation of domains containing different amounts of alpha-tocopherol. We suggest that, in these alpha-tocopherol-rich domains, the influence exerted by the vitamin on the phospholipids is strong enough to alter their thermotropic properties such that an additional endotherm appears in the thermogram, a characteristic not observed in homoacid phosphatidylcholines. alpha-Tocopherol caused a concentration-dependent removal of the detectable phase transition in all cases. The magnitude of the influence of alpha-tocopherol on phospholipid was dependent on the degree of unsaturation of the sn-2 acyl chain. These results are explained on the basis of the effect of alpha-tocopherol which will reduce the differences between gel and liquid crystalline states, the magnitude of these differences depending on the type of phospholipid considered, which are probably related to the change of molecular shape of phosphatidylcholines containing a polyunsaturated acyl chain.

The phase behavior of aqueous dispersions of unsaturated mixtures of diacylglycerols and phospholipids.

The phase behavior of mixtures of 1-palmitoyl-2-oleoyl-sn-glycerol (1,2-POG) with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS) was studied by using DSC, small-angle X-ray diffraction and 31P-NMR. The results have been used to construct phase diagrams for both type of mixtures, in the 0-45 degreesC range. It is concluded that 1, 2-POG form complexes in the gel phases with both POPC and POPS. In the case of POPC, two complexes are postulated, the first one at a 1, 2-POG/POPC molar ratio of 40:60, and the second one at 70:30, defining three different regions in the phase diagram. Two eutectic points are proposed to occur: one at a very low 1,2-POG concentration and the other at a 1,2-POG concentration slightly lower than 70%. In the case of the 1,2-POG/POPS mixtures, the pattern was similar, but the first complex was seen to happen at a higher concentration, about 50 mol% of 1,2-POG, whereas the second was found at 80 mol% of 1,2-POG. This indicated a bigger presence of 1,2-POG in the complexes with POPS than with POPC. In the first region of the phase diagram, i.e. at concentrations of 1,2-POG lower than that required for the formation of the first complex, and at temperatures above the phase transition, lamellar phases were seen in all the cases. In region 2 of the phase diagram, i.e. at concentrations where the first and the second complexes coexist, a mixture of lamellar and non-lamellar phases was observed. Finally, at high concentrations of 1,2-POG, non-lamellar phases were detected as predominant, these phases being of an isotropic nature, according to 31P-NMR. An important conclusion of this study is that, using unsaturated lipids, similar to those found in biological membranes, it has been shown that diacylglycerols are found separated in domains, and that this process starts at very low concentrations of diacylglycerols. The formation of separated domains enriched in diacylglycerol is biologically relevant as it will allow them to have important effects on the membrane structure besides the fact that their concentration in the biomembrane is relatively low.

Modulation of polymorphic properties of dielaidoylphosphatidylethanolamine by the antineoplastic ether lipid 1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine.

The capacity of the antineoplastic ether lipid 1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine (ET-18-OCH3) to modulate the polymorphic properties of dielaidoylphosphatidylethanolamine has been studied using biophysical techniques. Differential scanning calorimetry showed that ET-18-OCH3 depresses the onset of the Lbeta to Lalpha phase transition, decreasing also DeltaH of the transition. At the same time, the onset of the transition from Lalpha to inverted hexagonal HII phase was gradually increased as the ether lipid concentration was increased, totally disappearing at concentrations higher than 5 mol%. Small-angle X-ray diffraction and 31P-NMR confirmed that ET-18-OCH3 induced that the appearance of the inverted hexagonal HII phase was shifted towards higher temperatures completely disappearing at concentrations higher than 5 mol%. These results were used to elaborate a partial phase diagram and they were discussed as a function of the molecular action of ET-18-OCH3.

Fluorescence study of a derivatized diacylglycerol incorporated in model membranes.

A fluorescence study of a diacylglycerol derivatized with the n-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) chromophore (NBD) was carried out. Fluorescence self-quenching was observed for this probe in lecithin model membranes due to collisional interaction rather than to an aggregational behaviour of the probe. The efficient energy migration (Ro = 28 A) of the NBD fluorophore was studied via the fluorescence depolarization upon increase of probe concentration in membranes, and the results are compared with a model where a random distribution of the probes is assumed. A surface location of the chromophore was concluded for the NBD derivative of diacylglycerol, both from the fluorescence parameters and from the study of its fluorescence quenching by spin label probes. Very high lateral diffusion coefficients were obtained for these probes, both from the self-quenching (D = 2-6 x 10(-6) cm2 s-1) and from the spin probe quenching (D = 3.5 x 10(-6) cm2 s-1) studies. A concomitant fluorescence study of the related probe NBD-phosphatidylcholine revealed that its photophysical behaviour is similar to the derivatized diacylglycerol.

1,2-Dioleoylglycerol promotes calcium-induced fusion in phospholipid vesicles.

The effect of 1,2-dioleoyglycerol (1,2-DOG) on the promotion of Ca(2+)-induced fusion of phosphatidylserine/phosphatidylcholine (PS/PC) vesicles was studied. 1,2-DOG is able to induce the mixing of membrane lipids at concentrations of 10 mol% without mixing of vesicular contents. At concentrations of 20 mol% or higher, 1,2-DOG promotes fusion, lipid and content mixing, of LUV composed of an equimolar mixture of PS and PC, which otherwise are unable to fuse in the presence of Ca2+. Fusion was demonstrated by fluorescence assays monitoring mixing of aqueous vesicular contents and mixing of membrane lipids. Studies by Fourier transform infrared spectroscopy provided evidence for a fusion mechanism different to that of Ca(2+)-induced fusion of pure PS vesicles. Final equilibrium structures were characterized by 31P-NMR and freeze-fracture electron microscopy. Ca(2+)-induced fusion of 1,2-DOG containing vesicles is accompanied by the formation of isotropic structures which are shown to correspond to structures with lipidic particle morphology. The possible fusion mechanisms and implications are discussed.