1-2-dielaidoylphosphatidylethanolamine and 1-2-dioleoylphosphatidylserine

Desmin, an intermediate filament protein expressed in muscle cells, plays a key role in the integrity and regulation of the contractile system. Furthermore, the distribution of desmin in cells and its interplay with plasma and organelle membranes are crucial for cell functions; however, the fundamental properties of lipid-desmin interactions remain unknown. Using a water-in-oil method for a limited space system in vitro, we examined the distribution of desmin in three types of phospholipid droplets: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-dioleoyl-sn-glycero-3-phosphoserine (DOPS). When fluorescent-labeled desmin was observed for 60 min after desmin assembly was initiated by adding 25 mM KCl, desmin accumulated on both the DOPE and DOPS layers; however, it did not accumulate on the DOPC layer of droplets. An increase in salt concentration did not moderate the accumulation. The initial form of either oligomer or mature filament affected the accumulation on each lipid layer. When liposomes were included in the droplets, desmin was associated with DOPE but not on DOPC liposomes. These results suggest that desmin has the potential for association with phospholipids concerning desmin form and lipid shape. The behavior and composition of living membranes may affect the distribution of desmin networks.

Topics: Animals; Chickens; Desmin; Intermediate Filaments; Lipid Droplets; Liposomes; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Phospholipids

Parkinson's disease (PD), closely associated with the misfolding and aggregation of the neuronal protein α-synuclein (A-Syn), is a neurodegenerative disorder with no cure to date. Here, we show that the commercially available, inexpensive, aminoglycoside antibiotic kanamycin effectively inhibits both lipid-induced and solution-phase aggregation of A-Syn in vitro, pointing towards the prospective repurposing of kanamycin as a potential anti-PD drug.

Topics: alpha-Synuclein; Anti-Bacterial Agents; Cell Line, Tumor; Humans; Kanamycin; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Protein Binding; Protein Conformation; Protein Multimerization; Unilamellar Liposomes

Three-dimensional (3D) single-particle tracking was employed to study the lipid membrane morphology change at different pHs on glass supported lipid bilayers (SLBs) [1,2-dioleoyl- sn-glycero-3-phosphoethanolamine/1,2-dioleoyl- sn-glycero-3-phospho-l-serine (sodium salt)/1,2-dioleoyl- sn-glycero-3-phosphocholine = 5:3:2]. Fluorescently tagged, carboxylated polystyrene nanoparticles (of 100 nm) were used as the probes. At neutral pHs, the particles' diffusion was close to two-dimensional Brownian motion, indicating a mainly planar structure of the SLBs. When the environmental pH was tuned to be basic at 10.0, transiently confined diffusions within small areas were frequently observed. These confinements had a lateral dimension of 100-200 nm. Most interestingly, they showed 3D bulged structures protruding from the planar lipid bilayer. The particles were trapped by these 3D structures for a short period of time (∼0.75 s), with an estimated escape activation energy of ∼4.2 k

Topics: Diffusion; Hydrogen-Ion Concentration; Lipid Bilayers; Nanoparticles; Phosphatidylethanolamines; Phosphatidylserines; Polystyrenes; Surface Properties

In the present work we introduce a straightforward fluorescent assay that can be applied in studies of the transbilayer movement (flip-flop) of fluorescent lipid analogues across supported phospholipid bilayers (SPBs). The assay is based on the distance dependent fluorescence quenching by light absorbing surfaces. Applied to SPBs this effect leads to strong differences in fluorescence lifetimes when the dye moves from the outer lipid leaflet to the leaflet in contact with the support. Herein, we present the basic principles of this novel approach, and comment on its advantages over the commonly used methods for investigating flip-flop dynamics across lipid bilayers. We test the assay on the fluorescent lipid analog Atto633-DOPE and the 3-hydroxyflavone F2N12S probe in SPBs composed of DOPC/ DOPS lipids. Moreover, we compare and discuss the flip-flop rates of the probes with respect to their lateral diffusion coefficients.

Topics: Chemistry Techniques, Analytical; Diffusion; Fluorescence; Fluorescent Dyes; Kinetics; Lipid Bilayers; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Phospholipids; Time Factors

The membrane surface charge modifies the conductance of ion channels by changing the electric potential and redistributing the ionic composition in their vicinity. We have studied the effects of lipid charge on the conductance of a multi-state channel formed in planar lipid bilayers by the peptide antibiotic alamethicin. The channel conductance was measured in two lipids: in a neutral dioleoylphosphatidylethanolamine (DOPE) and a negatively charged dioleoylphosphatidylserine (DOPS). The charge state of DOPS was manipulated by the pH of the membrane-bathing solution. We find that at high salt concentrations (e.g., 2 M NaCl) the effect of the lipid charge is below the accuracy of our measurements. However, when the salt concentration in the membrane-bathing solution is decreased, the surface charge manifests itself as an increase in the conductance of the first two channel levels that correspond to the smallest conductive alamethicin aggregates. Our analysis shows that both the salt and pH dependence of the surface charge effect can be rationalized within the nonlinear Poisson-Boltzmann approach. Given channel conductance in neutral lipids, we use different procedures to account for the surface charge (e.g., introduce averaging over the channel aperture and take into account Na+ adsorption to DOPS heads), but only one adjustable parameter: an effective distance from the nearest lipid charge to the channel mouth center. We show that this distance varies by 0.3-0.4 nm upon channel transition from the minimal conducting aggregate (level L0) to the next larger one (level L1). This conclusion is in accord with a simple geometrical model of alamethicin aggregation.

Topics: Alamethicin; Anti-Bacterial Agents; Biophysical Phenomena; Biophysics; Electric Conductivity; Hydrogen-Ion Concentration; In Vitro Techniques; Ion Channels; Ionophores; Lipid Bilayers; Models, Chemical; Phosphatidylethanolamines; Phosphatidylserines; Sodium Chloride; Static Electricity

The cyclic lipodepsipeptide, syringomycin E, when incorporated into planar lipid bilayer membranes, forms two types of channels (small and large) that are different in conductance by a factor of sixfold. To discriminate between a cluster organization-type channel structure and other possible different structures for the two channel types, their ionic selectivity and pore size were determined. Pore size was assessed using water-soluble polymers. Ion selectivity was found to be essentially the same for both the small and large channels. Their reversal (zero current) potentials with the sign corresponding to anionic selectivity did not differ by more than 3 mV at a twofold electrolyte gradient across the bilayer. Reduction in the single-channel conductance induced by poly(ethylene glycol)s of different molecular weights demonstrated that the aqueous pore sizes of the small and large channels did not differ by more than 2% and were close to 1 nm. Based on their virtually identical selectivity and size, we conclude that large syringomycin E channels are clusters of small ones exhibiting synchronous opening and closing.

Topics: Anti-Bacterial Agents; Electric Conductivity; Electrolytes; Ion Channels; Lipid Bilayers; Membrane Potentials; Models, Biological; Peptides, Cyclic; Phosphatidylethanolamines; Phosphatidylserines

Phospholipase A2 (PLA2)-mediated hydrolysis of membrane phospholipids was measured by ellipsometry, and the inhibition of this process by annexin V was studied. Planar membranes, consisting of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine (PC/PE/PS; 54:33:13, on molar basis), were degraded by pancreatic PLA2, and the rate of hydrolysis was limited to about 0.7%/min. The influence of graded coverage of the membrane with annexin V was studied. The degree of PLA2 inhibition was nonlinearly related to the amount of membrane-bound annexin V, and binding of only 12% and 54% of full membrane coverage resulted in, respectively, 50% and 93% inhibition. These findings indicate that the inhibition of PLA2-mediated hydrolysis by annexin V cannot be simply explained by shielding of phospholipid substrates from the enzyme. Moreover, the present results leave room for a role of endogenous annexin V in regulating phospholipid turnover in the plasma membrane of parenchymal cells such as cardiomyocytes.

Topics: Animals; Annexin A5; Kinetics; Lipid Bilayers; Pancreas; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Phospholipases A; Phospholipases A2; Surface Properties; Swine

The ability of three sterols of different structure to influence the interaction of syringomycin E (an antifungal antibiotic that forms voltage dependent channels in planar lipid bilayers) with a planar lipid bilayer was evaluated. The rate of increase of bilayer conductance induced by syringomycin E was about 1000-times less in bilayers containing 50 mol% of cholesterol compared to bilayers without sterols. The effect of ergosterol (the primary sterol of fungal cells) on the sensitivity of bilayers to syringomycin E was much weaker than that of cholesterol, while stigmasterol (one of the main sterols of plant cells) did not significantly influence the ability of syringomycin E to induce a conductance increase in the bilayer. None of the sterols altered the single channel conductance properties of syringomycin E. These observations suggest that cholesterol affects the sensitivity of target membranes to syringomycin E by enlarging the energy barrier for channel formation rather than participating in channel formation itself.

Topics: Antifungal Agents; Bacterial Proteins; Cholestenes; Electric Conductivity; Ion Channels; Lipid Bilayers; Phosphatidylethanolamines; Phosphatidylserines

The response of coagulation tests to heparin can be expressed as the coagulation time of plasma containing heparin divided by the coagulation time of the same plasma without heparin (CT ratio). The purpose of the present study was to assess the influence of liposomes on these response to heparin of four coagulation tests: the kaolin-induced coagulation time, the tissue factor-induced coagulation time, the factor Xa-induced coagulation time, and the thrombin-induced coagulation time. Liposomes were prepared from dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylethanolamine (DOPE), and dioleoylphosphatidylserine (DOPS). High concentrations of DOPS/DOPE/DOPC (20:40:40) liposomes enhanced the CT ratio of the four coagulation tests, more than DOPS/DOPC (20:80) or DOPE/DOPC (40:60) or a mixture of DOPS/DOPC and DOPE/DOPC liposomes. These experiments demonstrate that there is synergism between DOPS and DOPE in promoting heparin's anticoagulant effect if both phospholipids are incorporated into the same liposome surface.

Topics: Blood Coagulation; Blood Coagulation Tests; Drug Synergism; Heparin; Humans; Kaolin; Liposomes; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Thromboplastin

Antioxidant reactions of mixtures of vitamin E, vitamin C and phospholipids in autoxidizing lipids at 90 degrees C have been studied by ESR spectroscopy. When the phospholipid contained a tertiary amine (e.g. phosphatidylcholine), the vitamin C and the vitamin E radicals were successively observed as these two vitamins were sequentially oxidised during lipid oxidation. In the presence of the primary amine contained in phosphatidylserine, the vitamin E oxidation was delayed for a few hours. In this case neither the vitamin C, nor the vitamin E radicals but a nitroxide radical derived from the phospholipid was observed. Similar results to those obtained with PS were obtained in the presence of either phosphatidylethanolamine or soybean lecithin. The participation in the radical reactions of phospholipids possessing a primary amine can therefore explain the synergistic effect of these phospholipids in a mixture of vitamins E and C.

Topics: Ascorbic Acid; Electron Spin Resonance Spectroscopy; Fatty Acids, Unsaturated; Free Radicals; Hot Temperature; Oxidation-Reduction; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylinositols; Phosphatidylserines; Phospholipids; Vitamin E

Adhesion between patch-clamped lipid membranes and glass micropipettes is measured by high contrast video imaging of the mechanical response to the application of suction pressure across the patch. The free patch of membrane reversibly alters both its contact angle and radius of curvature on pressure changes. The assumption that an adhesive force between the membrane and the pipette can sustain normal tension up to a maximum Ta at the edge of the free patch accounts for the observed mechanical responses. When the normal component of the pressure-induced membrane tension exceeds Ta membrane at the contact point between the free patch and the lipid-glass interface is pulled away from the pipette wall, resulting in a decreased radius of curvature for the patch and an increased contact angle. Measurements of the membrane radius of curvature as a function of the suction pressure and pipette radius determine line adhesion tensions Ta which range from 0.5 to 4.0 dyn/cm. Similar behavior of patch-clamped cell membranes implies similar adhesion mechanics.

Topics: Adhesiveness; Cholesterol; Glass; Hydrostatic Pressure; Lipid Bilayers; Mathematics; Models, Theoretical; Phosphatidylethanolamines; Phosphatidylserines

We investigated the effect of the antineoplastic drug doxorubicin on the order of the acyl chains in liquid-crystalline mixed bilayers consisting of dioleoylphosphatidylserine (DOPS) or -phosphatidic acid (DOPA), and dioleoylphosphatidylcholine (DOPC) or -phosphatidylethanolamine (DOPE). Previous 2H-NMR studies on bilayers consisting of a single species of di[11,11-2H2]oleoyl-labeled phospholipid showed that doxorubicin does not affect the acyl chain order of pure zwitterionic phospholipid but dramatically decreases the order of anionic phospholipid [de Wolf, F. A., et al. (1991) Biochim. Biophys. Acta 1096, 67-80]. In the present work, we studied mixed bilayers in which alternatively the anionic or the zwitterionic phospholipid component was 2H-labeled so as to monitor its individual acyl chain order. Doxorubicin decreased the order parameter of the mixed anionic and zwitterionic lipids by approximately the same amount and did not induce a clear segregation of the lipid components into extended, separate domains. The drug had a comparable disordering effect on mixed bilayers of unlabeled cardiolipin and 2H-labeled zwitterionic phospholipid, indicating the absence of extensive segregation also in that case. Upon addition of doxorubicin to bilayers consisting of 67 mol% DOPE and 33 mol% anionic phospholipid, a significant part of the lipid adopted the inverted hexagonal (HII) phase at 25 degrees C. This bilayer destabilization, which occurred only in mixtures of anionic phospholipid and sufficient amounts of DOPE, might be of physiological importance. Even upon formation of extended HII-phase domains, lipid segregation was not clearly detectable, since the relative distribution of 2H-labeled anionic phospholipid and [2H]DOPE between the bilayer phase and HII phase was very similar. Our findings argue against a role of extensive anionic/zwitterionic lipid segregation in the mechanism of action and toxicity of doxorubicin.

Topics: Doxorubicin; Membranes, Artificial; Models, Biological; Molecular Structure; Phosphatidic Acids; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Phospholipids; Structure-Activity Relationship

The interaction of phenethyl alcohol with model membranes and its effect on translocation of the chemically prepared mitochondrial precursor protein apocytochrome c across a lipid bilayer was studied. Phenethyl alcohol efficiently penetrates into monolayers and causes acyl chain disordering judged from deuterium nuclear magnetic resonance measurements with specific acyl chain-deuterated phospholipids. Translocation of apocytochrome c across a phospholipid bilayer was stimulated on addition of phenethyl alcohol indicating that the efficiency of translocation of this precursor protein is enhanced due to a disorder of the acyl chain region of the bilayer.

Topics: Apoproteins; Biological Transport; Cytochrome c Group; Cytochromes c; Ethanol; Lipid Bilayers; Magnetic Resonance Spectroscopy; Membrane Lipids; Mitochondria; Phenylethyl Alcohol; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylserines; Phospholipids; Protein Precursors; Trypsin