RESUMO
We studied coexisting micro- and nanoscopic liquid-ordered/liquid-disordered domains in fully hydrated multilamellar vesicles using small-angle X-ray scattering. Large domains exhibited long-range out-of-plane positional correlations of like domains, consistent with previous reports. In contrast, such correlations were absent in nanoscopic domains. Advancing a global analysis of the in situ data allowed us to gain a deep insight into the structural and elastic properties of the coexisting domains, including the partitioning of cholesterol in each domain. In agreement with a previous report, we found that the thickness mismatch between ordered and disordered domains decreased for nanoscopic domains. At the same time, we found also the lipid packing mismatch to be decreased for nano-domains, mainly due to the liquid-disordered domains becoming more densely packed when decreasing their size.
RESUMO
We have studied the electrostatic screening effect of NaCl solutions on the interactions between anionic lipid bilayers in the fluid lamellar phase using a Poisson-Boltzmann-based mean-field approach with constant charge and constant potential limiting charge regulation boundary conditions. The full DLVO potential, including the electrostatic, hydration and van der Waals interactions, was coupled to thermal bending fluctuations of the membranes via a variational Gaussian Ansatz. This allowed us to analyze the coupling between the osmotic pressure and the fluctuation amplitudes and compare them both simultaneously with their measured dependence on the bilayer separation, determined by the small-angle X-ray scattering experiments. High-structural resolution analysis of the scattering data revealed no significant changes of membrane structure as a function of salt concentration. Parsimonious description of our results is consistent with the constant charge limit of the general charge regulation phenomenology, with fully dissociated lipid charge groups, together with a 6-fold reduction of the membranes' bending rigidity upon increasing NaCl concentration.
Assuntos
Elasticidade , Bicamadas Lipídicas/química , Ânions , Pressão Osmótica , Eletricidade EstáticaRESUMO
Specular neutron reflectometry was measured on a floating bilayer system consisting of 1,2-dipalmitoyl-d62-sn-glycero-3-phosphocholine deposited over a 1,2-dibehenoyl-sn-glycero-3-phosphocholine bilayer at 25 and 55 °C. The internal structure of lipid bilayers was described by a one-dimensional neutron scattering length density profile model, originally developed for the evaluation of small-angle scattering data. The reflectivity data from the supported bilayer were evaluated separately and used further as constraints in modeling the floating bilayer reflectivity curves. The model reflectivity curves successfully describe the experimental reflectivities of the supported bilayer in the gel phase and the floating bilayer system in the liquid-crystalline phase. The results yield an internal structure of a deposited bilayer and a floating bilayer on the level of component groups of lipid molecules. The obtained structure of the floating d62-diC16:0PC bilayer displays high resemblance of the bilayer structure in the form of unilamellar vesicles. At the same time, however, the results show differences in comparison to unilamellar vesicle bilayers, most likely due to the undulations of supported bilayers.
Assuntos
Dimiristoilfosfatidilcolina/química , Bicamadas Lipídicas/química , Fosfatidilcolinas/química , Lipossomas Unilamelares/química , Difração de Nêutrons , Espalhamento a Baixo ÂnguloRESUMO
Small-angle neutron scattering data were collected from aqueous dispersions of unilamellar vesicles (ULVs) consisting of mixtures of 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine and a homologous series of N,N-dimethyl-N-alkylamine-N-oxides (CnNO, n = 12, 14, 16, and 18, where n is the number of carbon atoms in the alkyl chain). A modeling approach was applied to the neutron scattering curves to obtain the bilayer structural parameters. Particularly, the external (2)H2O/H2O contrast variation technique was carried out on pure dioleoylphosphatidylcholine (DOPC) ULVs to determine the hydrophilic region thickness [Formula: see text] = 9.8 ± 0.6 Å. Consequently, the hydrocarbon region thickness [Formula: see text], the lateral bilayer area per one lipid molecule [Formula: see text], and the number of water molecules located in the hydrophilic region per one lipid molecule [Formula: see text] were obtained from single-contrast neutron scattering curves using the previously determined [Formula: see text]. The structural parameters were extracted as functions of [Formula: see text] (the CnNO:DOPC molar ratio) and n. The dependences [Formula: see text] provided the partial lateral areas of CnNOs ([Formula: see text]) and DOPC ([Formula: see text]) in bilayers. It was observed that the [Formula: see text]'s were constant in the investigated interval of [Formula: see text] and for n = 12, 14, and 16 equal to 36.6 ± 0.4 Å(2), while [Formula: see text] increased to 39.4 ± 0.4 Å(2). The bilayer hydrocarbon region thickness [Formula: see text] decreased with intercalation of each CnNO. This effect increased with [Formula: see text] and decreased with increasing CnNO alkyl chain length. The intercalation of C18NO changed the [Formula: see text] only slightly. To quantify the effect of CnNO intercalation into DOPC bilayers we fit the [Formula: see text] dependences with weighted linear approximations and acquired their slopes [Formula: see text].
Assuntos
Dimetilaminas/farmacologia , Bicamadas Lipídicas/química , Difração de Nêutrons , Fosfatidilcolinas , Espalhamento a Baixo Ângulo , Lipossomas Unilamelares/química , Relação Dose-Resposta a DrogaRESUMO
The volumetric properties of fluid bilayers formed of dioleoylphosphatidylcholine (DOPC) with incorporated N,N-dimethyl-N-alkylamine N-oxides (CnNO, n=6, 10-18 is the even number of carbons in alkyl chain) were studied by vibrating tube densitometry in the temperature interval from 20°C to 50°C. It was found that the DOPC and CnNO mixed ideally in the investigated composition range and hence the molecular volumes of DOPC (VDOPC) and incorporated CnNO (VCnNO) were constant and additive within error limits. From the temperature dependencies of the molecular volumes of DOPC and CnNO their coefficients of isobaric thermal expansivities in the investigated temperature interval were obtained. The VCnNO volumes of CnNO incorporated into DOPC bilayers showed linear dependencies on the CnNO alkyl chain length at all measured temperatures. This allowed to calculate the component volume of the CnNO methylene group (VCH2) at several temperatures and its coefficient of isobaric thermal expansivity. Using the assumption that the component volume of the CnNO methyl group VCH3 = 2VCH2 we also calculated the component volume and the coefficient of isobaric thermal expansivity of the hydrophilic group of CnNO (VNO). We found that the VCH2 volume increases in the whole temperature interval whereas the VNO volume decreases.