1-palmitoyl-2-oleoylphosphatidylcholine and 1-2-dilinoleoyl-3-phosphatidylethanolamine

Packing defects in lipid bilayer play a significant role in the biological activities of cell membranes. Time-resolved fluorescence depolarization has been used to detect and characterize the onset of packing defects in binary mixtures of dilinoleoylphosphatidylethanolamine/1-palmitoyl-2- oleoylphosphatidylcholine (PE/PC). These PE/PC mixtures exhibit mesoscopic packing defect state (D), as well as one-dimensional lambellar liquid crystalline (L alpha) and two-dimensional inverted hexagonal (HII) ordered phases. Based on previous electron microscopic investigations, this D state is characterized by the presence of interlamellar attachments and precursors of HII phase between the lipid layers. Using a rotational diffusion model for rod-shaped fluorophore in a curved matrix, rotational dynamics parameters, second rank order parameter, localized wobbling diffusion, and curvature-dependent rotational diffusion constants of dipyenylhexatriene (DPH)-labeled PC (DPH-PC) in the host PE/PC matrix were recovered from the measured fluorescence depolarization decays of DPH fluorescence. At approximately 60% PE, abrupt increases in these rotational dynamics parameters were observed, reflecting the onset of packing defects in the host PE/PC matrix. We have demonstrated that rotational dynamics parameters are very sensitive in detecting the onset of curvature-associating packing defects in lipid membranes. In addition, the presence of the D state can be characterized by the enhanced wobbling diffusional motion and order packing of lipid molecules, and by the presence of localized curvatures in the lipid layers.

Topics: Cell Membrane; Diffusion; Kinetics; Lipid Bilayers; Mathematics; Models, Biological; Phosphatidylcholines; Phosphatidylethanolamines; Spectrometry, Fluorescence; Time Factors

Bilayer packing defects in binary dilinoleoylphosphatidylethanolamine and 1-palmitoyl-2-oleoylphosphatidylcholine (DLPE/POPC) lipid mixtures have been studied by the use of nanosecond-resolved intramolecular excimer fluorescence spectroscopy. Frequency-domain fluorescence intensity decays of dual-chain labelled dipyrenyl lipids of different chain lengths in DLPE/POPC mixtures were acquired at both the monomer (392 nm) and excimer (475 nm) emission channels and at 20 degrees C. On the basis of a new intramolecular excimer formation kinetic model, the extent of aggregation and the rotational mobility, in terms of the equilibrium constant of the monomer to aggregated state and the excimer association rate constant, respectively, of the intralipid pyrenes were calculated from the frequency-domain data. Within the range of 60-100% DLPE where bilayer defects are known to coexist with bilayer and non-bilayer states, a prominent peak in the equilibrium constant and a concomitant dip in the excimer association constant at approximately 80% DLPE were observed. Our nanosecond-resolved fluorescence results suggest that the intramolecular excimer kinetic parameters of dipyrenyl lipids are very sensitive to the onset of bilayer packing defects in lipid membranes. Moreover, the onset of bilayer defect state is characterized by the greater extent of aggregation and more hindered rotational mobility of the acyl chains as compared with the bilayer (0% DLPE) and non-bilayer inverted hexagonal (100% DLPE) states of the lipid membranes.

Topics: Kinetics; Lipid Bilayers; Models, Chemical; Molecular Conformation; Phosphatidylcholines; Phosphatidylethanolamines; Spectrometry, Fluorescence

The bilayer stability of liposomes containing unsaturated phosphatidylethanolamine (PE) has been investigated by measuring the C = O and CH2 stretching frequencies of the lipids at different lipid compositions and temperatures. Binary mixtures of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) and dilinoleoyl-PE (DLPE) are known to exhibit lamellar liquid crystalline (L alpha), inverted hexagonal (HII) and metastable intermediate (I) phases. Abrupt increases in the C = O and CH2 stretching frequencies at 65-75 and 90-95% PE, respectively, were found as the PE content of the DLPE/POPC mixtures was increased from 0 to 100%. These transitions were associated with the L alpha -I and I-HII phase transitions of the DLPE/POPC mixtures, accordingly. The effects of three lipid perturbants, butylated hydroxytoluene (BHT), diacylglycerol (DG) and cholesterol (CL), on the above L alpha -I and I-HII transitions were also examined. All perturbants were found to be effective in shifting the L alpha -I transition of the DLPC/POPC mixtures to a lower PE% as detected by the C = O stretching frequency measurements. On the other hand, the perturbants appeared to eliminate the I-HII transition of the DLPE/POPC mixtures as detected by the CH2 stretching frequency measurements. The effectiveness of the perturbants in promoting the L alpha -I phase transition of the DLPE/POPC mixtures followed the order of DG > BHT > CL.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Lipid Bilayers; Phosphatidylcholines; Phosphatidylethanolamines; Spectrophotometry, Infrared

The hydrolysis of mixed dilinoleoylphosphatidylethanolamine (DiLinPE) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) dispersions by porcine phospholipase A2, under conditions leading to the bilayer-to-nonbilayer phase transition, has been studied. Two structurally distinct forms of the dispersions were used, multilamellar vesicles (MLV) and supercritical large unilamellar vesicles (SCLUV). In MLV, maximum free fatty acid was produced in dispersions containing 85 mol % DiLinPE. The peak in the fatty acid release is found at the onset of appearance of the nonbilayer defects reported earlier. DiLinPE was found to be preferentially hydrolyzed as compared to POPC. When cholesterol was added to the mixed DiLinPE/POPC MLV, the onset of the observable appearance of nonbilayer defects, the positions of the peaks for total hydrolysis, and the preferential hydrolysis of DiLinPE were all shifted toward lower DiLinPE concentrations. In SCLUV, where the appearance of nonbilayer structures is prevented by constraining the lipids in bilayer configuration, the hydrolysis by PLA2 increases with increasing DiLinPE as predicted from the increase in the calculated monolayer bending energy. The results are interpreted to be related to the pretransition molecular-packing stress and defects at the onset of the bilayer-to-nonbilayer transition. Results indicate that the porcine pancreatic phospholipase A2 activity is controlled by bilayer-packing stress, which may cause structural defects of the substrate, among other factors. Results also indicate a preferential localization of PE at stress-related defect regions.

Topics: Animals; Cholesterol; Enzyme Activation; Freeze Fracturing; Hydrolysis; Kinetics; Lipid Bilayers; Phosphatidylcholines; Phosphatidylethanolamines; Phospholipases A; Phospholipases A2; Swine

The polymorphic phase behaviour of dilinoleoylphosphatidyethanolamine (DLPE) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) is investigated by freeze-fracture electron microscopy, X-ray diffraction and 31P-NMR. The structures at 5% or less POPC are predominantly inverted hexagonal (HII), whereas at 15% or more POPC, the structure is mostly bilayer (L), interrupted by defects (lipidic particles). A cubic phase structure is observed in the transition range between H and L phases; the cubic arrangement deteriorates at higher temperatures into an amorphous aggregate of spherical units. Both cubic and amorphous structures contribute to the isotropic 31P resonance, with no preference for PC or PE partitioning in the isotropic motion as observed by high resolution NMR. The existence of the cubic phase seems to depend cirtically on the homogeneity and the degree unsaturation of the phospholipids.

Topics: Freeze Fracturing; Lipid Bilayers; Liposomes; Magnetic Resonance Spectroscopy; Microscopy, Electron; Models, Biological; Molecular Conformation; Phosphatidylcholines; Phosphatidylethanolamines; X-Ray Diffraction