1-palmitoyl-2-oleoylphosphatidylcholine and dodecyloctaethyleneglycol-monoether

The cationic large unilamellar mixed liposomes from 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) and didodecyldimethylammonium bromide (DDAB) or dioctadecyldimethylammonium bromide (DODAB) were prepared. The influence of the addition of Triton X-100 (TX-100) or octaethylene glycol mono-n-dodecylether (C(12)E(8)) on the membrane integrity was investigated turbidimetrically. The stability of the liposomal systems was estimated by monitoring fluorimetrically at 25 °C the rate of spontaneous and surfactant-induced release of entrapped 5(6)-carboxyfluorescein (CF). In order to evaluate the interaction of the cationic DODAB guest with the host POPC membrane, the main phase transition temperatures (T(m)) were determined by electron paramagnetic resonance spectroscopy (EPR). All the results obtained show that the presence of DODAB and DDAB stabilizes the POPC liposomes. The extent of stabilization depends on the concentration and nature of the cationic guest.

Topics: Cations; Electron Spin Resonance Spectroscopy; Kinetics; Lipid Bilayers; Liposomes; Molecular Dynamics Simulation; Nephelometry and Turbidimetry; Octoxynol; Phase Transition; Phosphatidylcholines; Polyethylene Glycols; Quaternary Ammonium Compounds; Transition Temperature

We have studied the effect of two cosolvents, urea and glycerol, on the association and interactions of a surfactant, octaethyleneglycol dodecyl ether (C(12)EO(8)) and a phospholipid (POPC). We have measured the CMC, the partition coefficient, the effective mole fractions X(e)(sat) and X(e)(sol) at the onset and completion of the membrane-to-micelle transition (membrane solubilization), and the enthalpies of transfer of surfactant by ITC. Changes in membrane order and dynamics were characterized by time-resolved fluorescence anisotropy measurements of DPH, and micelle sizes and clouding by light scattering. The cosolvents have complex effects that are not governed by the well-known 'salting in' or 'salting out' effects on the solubility alone. Instead, urea and glycerol alter also the intrinsic curvature ('effective molecular shape') of the detergent and the order and packing of the membrane. These curvature effects have an unusual enthalpy/entropy balance and are not additive for lipid and detergent. The results shed light on the phenomena governing lipid-detergent systems in general and have a number of implications for the use of cosolvents in membrane protein studies.

Topics: Calorimetry; Diphenylhexatriene; Fluorescent Dyes; Glycerol; Liposomes; Micelles; Phosphatidylcholines; Polyethylene Glycols; Solubility; Spectrometry, Fluorescence; Surface-Active Agents; Urea

Giant unilamellar phospholipid vesicles were prepared by the method of electroformation from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC). We studied the influence of different concentrations of the surfactant octaethyleneglycol dodecylether (C(12)E(8)) on the spontaneous shape transformations of POPC vesicles at room temperature. In accordance with previous results, we observed that low concentration of C(12)E(8) increased the speed of the characteristic vesicle shape transformation, starting from the initial shape with thin tubular protrusion, through beaded protrusion where the number of beads gradually decreased, to final spherical shapes with invagination, whereby the average mean curvature of the vesicle membrane monotonously decreased. In contrast, higher concentration of C(12)E(8) initially induced the shape transformation in the "opposite direction": in the protrusion, the number of beads gradually increased and eventually a tube was formed whereby the average mean curvature of the vesicle membrane gradually increased. However, at a certain point, an abrupt shape change took place to yield the vesicle with invagination. In this transition, the average mean curvature of the vesicle membrane discontinuously decreased. After this transition, the vesicle began to shrink and finally disappeared. We discuss possible mechanisms involved in the observed transformations.

Topics: Electrochemistry; Liposomes; Membrane Fluidity; Molecular Conformation; Phase Transition; Phosphatidylcholines; Phospholipids; Polyethylene Glycols; Solutions

We studied spontaneous shape transformations and burst of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) vesicles with exogeneously added non-ionic detergent octaethylene-glycol dodecylether C(12)E(8). The addition of C(12)E(8) increased the speed of the vesicle shape transformation, so that we were able to study for the first time the complete sequence of POPC vesicle shapes starting from initial spherical vesicle with long thin tubular protrusion to final shape with invagination(s). The average mean curvature of the vesicle membrane continuously decreases during this process. The shape of the invaginations is usually spherical, however also non-spherical shapes of invaginations were observed. C(12)E(8) increases amplitudes of the fluctuations of the vesicle membrane. At higher concentrations in the membrane, C(12)E(8) induces the membrane leakage and burst of the vesicles.

Topics: Algorithms; Detergents; Elasticity; Lipid Bilayers; Liposomes; Macromolecular Substances; Membrane Fluidity; Microscopy, Phase-Contrast; Models, Chemical; Molecular Conformation; Particle Size; Permeability; Phosphatidylcholines; Polyethylene Glycols; Surface Properties; Thermodynamics

The effects of a nonionic surfactant, octaethyleneglycol mono n-dodecyl ether (C12E8), on the electroporation of planar bilayer lipid membranes made of the synthetic lipid 1-pamitoyl 2-oleoyl phosphatidylcholine (POPC), was studied. High-amplitude ( approximately 100-450 mV) rectangular voltage pulses were used to electroporate the bilayers, followed by a prolonged, low-amplitude ( approximately 65 mV) voltage clamp to monitor the ensuing changes in transmembrane conductance. The electroporation thresholds of the membranes were found for rectangular voltage pulses of given durations. The strength-duration relationship was determined over a range from 10 micros to 10 s. The addition of C12E8 at concentrations of 0.1, 1, and 10 microM to the bath surrounding the membranes decreased the electroporation threshold monotonically with concentration for all durations (p < 0.0001). The decrease from control values ranged from 10% to 40%, depending on surfactant concentration and pulse duration. For a 10-micros pulse, the transmembrane conductance 150 micros after electroporation (G150) increased monotonically with the surfactant concentration (p = 0.007 for 10 microM C12E8). These findings suggest that C12E8 incorporates into POPC bilayers, allowing electroporation at lower intensities and/or shorter durations, and demonstrate that surfactants can be used to manipulate the electroporation threshold of lipid bilayers.

Topics: Detergents; Electric Conductivity; Electrochemistry; Electroporation; Kinetics; Lipid Bilayers; Models, Chemical; Phosphatidylcholines; Polyethylene Glycols; Surface-Active Agents; Time Factors

The interaction of the nonionic detergent octyl-beta-D-glucopyranoside (OG) with lipid bilayers was studied with high-sensitivity isothermal titration calorimetry (ITC) and solid-state 2H-NMR spectroscopy. The transfer of OG from the aqueous phase to lipid bilayers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) can be investigated by employing detergent at concentrations below the critical micellar concentration; it can be defined by a surface partition equilibrium with a partition coefficient of K = 120 +/- 10 M-1, a molar binding enthalpy of delta H degrees D = 1.3 +/- 0.15 kcal/mol, and a free energy of binding of delta G degrees D = -5.2 kcal/mol. The heat of transfer is temperature dependent, with a molar heat capacity of delta CP = -75 cal K-1 mol-1. The large heat capacity and the near-zero delta H are typical for a hydrophobic binding equilibrium. The partition constant K decreased to approximately 100 M-1 for POPC membranes mixed with either negatively charged lipids or cholesterol, but was independent of membrane curvature. In contrast, a much larger variation was observed in the partition enthalpy. delta H degrees D increased by about 50% for large vesicles and by 75% for membranes containing 50 mol% cholesterol. Structural changes in the lipid bilayer were investigated with solid-state 2H-NMR. POPC was selectively deuterated at the headgroup segments and at different positions of the fatty acyl chains, and the measurement of the quadrupolar splittings provided information on the conformation and the order of the bilayer membrane. Addition of OG had almost no influence on the lipid headgroup region, even at concentrations close to bilayer disruption. In contrast, the fluctuations of fatty acyl chain segments located in the inner part of the bilayer increased strongly with increasing OG concentration. The 2H-NMR results demonstrate that the headgroup region is the most stable structural element of the lipid membrane, remaining intact until the disordering of the chains reaches a critical limit. The perturbing effect of OG is thus different from that of another nonionic detergent, octaethyleneglycol mono-n-dodecylether (C12E8), which produces a general disordering at all levels of the lipid bilayer. The OG-POPC interaction was also investigated with POPC monolayers, using a Langmuir trough. In the absence of lipid, the measurement of the Gibbs adsorption isotherm for pure OG solutions yielded an OG surface area of AS = 51 +/-

Topics: Calorimetry; Chemical Phenomena; Chemistry, Physical; Cholesterol; Deuterium; Glucosides; Lipid Bilayers; Liposomes; Magnetic Resonance Spectroscopy; Models, Molecular; Phosphatidylcholines; Polyethylene Glycols; Temperature; Thermodynamics