We present a method for analyzing small angle x-ray scattering data on multilamellar phospholipid bilayer systems at full hydration. The method utilizes a modified Caillé theory structure factor in combination with a Gaussian model representation of the electron density profile such that it accounts also for the diffuse scattering between Bragg peaks. Thus the method can retrieve structural information even if only a few orders of diffraction are observed. We further introduce a procedure to derive fundamental parameters, such as area per lipid, membrane thickness, and number of water molecules per lipid, directly from the electron density profile without the need of additional volumetric measurements. The theoretical apparatus is applied to experimental data on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, and 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine liposome preparations.
Monolayer spontaneous curvatures for cholesterol, DOPE, POPE, DOPC, DPPC, DSPC, POPC, SOPC, and egg sphingomyelin were obtained using small-angle X-ray scattering (SAXS) on inverted hexagonal phases (HII). Spontaneous curvatures of bilayer forming lipids were estimated by adding controlled amounts to a HII forming template following previously established protocols. Spontaneous curvatures of both phosphatidylethanolamines and cholesterol were found to be at least a factor of two more negative than those of phosphatidylcholines, whose J0 values are closer to zero. Interestingly, a significant positive J0 value was retrieved for DPPC. We further determined the temperature dependence of the spontaneous curvatures J0(T) in the range from 15 to 55 °C, resulting in a quite narrow distribution of −1 to −3 × 10−3 (nm °C)−1 for most investigated lipids. The data allowed us to estimate the monolayer spontaneous curvatures of ternary lipid mixtures showing liquid ordered/liquid disordered phase coexistence. We report spontaneous curvature phase diagrams for DSPC/DOPC/Chol, DPPC/DOPC/Chol and SM/POPC/Chol and discuss effects on protein insertion and line tension.
We studied the influence of sodium and calcium chloride on the global and local membrane properties of fluid palmitoyl-oleoyl phosphatidylcholine bilayers, applying synchrotron small-angle x-ray diffraction, spin-labeling electron paramagnetic resonance spectroscopy, and differential scanning calorimetry, as well as simultaneous density and acoustic measurements. The salt concentration was varied over a wide range from 0 to 5 M. We found that NaCl leads to a continuous swelling of the bilayers, whereas the behavior of the bilayer separation dW in the presence of CaCl2 is more complex, showing an initial large dW value, which decreased upon further addition of salt and finally increased again in the high concentration regime. This can be understood by a change of balance between electrostatic and van der Waals interactions. We were further able to show that both salts lead to a significant increase of order within the lipid bilayer, leading to a decrease of bilayer elasticity and shift of main phase transition temperature. This effect is more pronounced for Ca2+, and occurs mainly in the high salt-concentration regime. Thus, we were able to reconcile previous controversies between molecular dynamics simulations and x-ray diffraction experiments regarding the effect of salts on neutral lipid bilayers.
For the first time the electron density of the lamellar liquid crystalline as well as of the inverted hexagonal phase could be retrieved at the transition temperature. A reliable decomposition of the d-spacings into hydrophobic and hydrophilic structure elements could be performed owing to the presence of a sufficient number of reflections. While the hydrocarbon chain length, d(C), in the lamellar phase with a value of 14.5 A lies within the extreme limits of the estimated chain length of the inverse hexagonal phase 10 A < d(C) < 16 A, the changes in the hydrophilic region vary strongly. During the lamellar-to-inverse hexagonal phase transition the area per lipid molecule reduces by approximately 25%, and the number of water molecules per lipid increases from 14 to 18. On the basis of the analysis of the structural components of each phase, the interface between the coexisting mesophases between 66 and 84 degrees C has been examined in detail, and a model for the formation of the first rods in the matrix of the lamellar phospholipid stack is discussed. Judging from the structural relations between the inverse hexagonal and the lamellar phase, we suggest a cooperative chain reaction of rod formation at the transition midpoint, which is mainly driven by minimizing the interstitial region.
The double-focusing high-flux wiggler beamline dedicated to small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) at ELETTRA has gone into user operation recently. It has been designed specifically for time-resolved studies of non-crystalline and fibrous materials in the submillisecond time scale, and has been optimized for small-angle scattering measurements. An overview of the beamline status and of some representative results, highlighting the performance of the SAXS beamline, are given.
The applicability of full‐q‐range models to fit low‐resolution X‐ray diffraction data from multibilayers exhibiting only weak quasi‐Bragg peak scattering has been analysed. The models consider different structure factors, accounting for different types of lattice disorder caused by stacking faults or bending fluctuations. Numerical tests of the models, considering instrumental influence of a line‐focus collimation system, demonstrated that Bragg peak line shapes given by different lattice disorders cannot be discerned. However, line‐shape parameters can be determined for a particular sample, if the type of disorder is known a priori. This has been verified by comparing the experimental results for the fluctuation parameter of palmitoyl‐oleoyl phosphatidylcholine (POPC) as a function of temperature with high‐resolution data on the same lipid. Tests further show that the calculation of structural parameters, such as the membrane thickness or the extent of the interbilayer water region, is not obscured by the smearing imposed by the instrument. The model was further applied successfully to experimental data of lipid mixtures composed of sphingomyelin (SM)/POPC/cholesterol and dipalmitoyl phosphatidylethanolamine (DPPE)/dipalmitoyl phosphatidylglycerol (DPPG). The structural parameters determined give valuable insight into the physical state of the membrane system, which is not accessible when quasi‐Bragg reflections only are considered.
Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. For the publisher's version, please access the DOI link below./ Pour consulter la version de l'éditeur, utilisez le lien DOI ci-dessous. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=f250b478-5ff0-4b72-8aa4-47a3c768b495 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=f250b478-5ff0-4b72-8aa4-47a3c768b495 Sciences, Schmiedlstrasse 6, 8042 Graz, Austria, and Raman Research Institute, Bangalore 560 080, India Received June 10, 2002. In Final Form: October 16, 2002 We have carried out X-ray and neutron diffraction experiments, as a function of temperature, on fully hydrated samples of dimyristoyl phosphatidylcholine bilayers. The data show the following: (a) In the vicinity of the LR to P ′ transition, we find an anomalous expansion of the water layer of ∼1.7 Å. (b) The lipid bilayer thickness increases quasi-linearly and is similar to the increase in the lamellar repeat spacing d found in dimyristoyl ethanolamine bilayers in the temperature range of TM to TM + 13°C. (c) In contrast to an earlier study, we find no significant changes to the steric size of the phosphatidylcholine headgroup. The anomalous increase in d is thus dominated by an expansion of the water layer. This expansion is caused by a distinct increase in bilayer fluctuations as revealed by an analysis of the Caillé parameter. Additional osmotic pressure experiments not only support this notion but have allowed us to further estimate the temperature dependence of both the bilayer bending rigidity, Kc, and the interbilayer compressional parameter, B. Both Kc and B experience an abrupt decrease on approaching TM from above, indicative of a "softening" of the bilayers. Structure and Interactions in the Anomalous
We have studied the concentration and temperature dependent influence of cholesterol, stigmasterol, and sitosterol on the global structure and the bending fluctuations of fluid dimyristoyl phosphatidylcholine and palmitoyl oleoyl phosphatidylcholine bilayers applying small-angle x-ray scattering, as well as dilatometry and ultrasound velocimetry. Independent of the lipid matrix, cholesterol was found to be most efficient in modulating bilayer thickness and elasticity, followed by sitosterol and stigmasterol. This can be attributed to the additional ethyl groups and double bond at the C(17) alkyl side-chain of the two plant sterols. Hence, it seems that some flexibility of the sterol hydrocarbon chain is needed to accommodate within the lipid bilayer. In addition, we did not observe two populations of membranes within the putative liquid-ordered/liquid-disordered phase coexistence regime of binary sterol/lipid mixtures. Instead, the diffraction patterns could be interpreted in terms of a uniform phase. This lends further support to the idea of compositional fluctuations of unstable sterol rich domains recently brought up by fluorescence microscopy experiments, which contrasts the formation of stable domains within the miscibility gap of binary lipid/sterol mixtures.
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