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Michael Rappolt

Publications and source records attributed to Michael Rappolt.

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Global SAXS data analysis for multilamellar vesicles: Evolution of the scattering density profile (SDP) model

We adapted the highly successful scattering density profile (SDP) model, which is used to jointly analyze small angle x-ray and neutron scattering data from unilamellar vesicles, for use with data from fully hydrated, liquid crystalline multilamellar vesicles (MLVs). By using a genetic algorithm, this new method is capable of providing high resolution structural information, as well as determining bilayer elastic bending fluctuations from standalone x-ray data. Important structural parameters, such as bilayer thickness and area per lipid were determined for a series of saturated and unsaturated lipids, as well as binary mixtures with cholesterol. Results are in good agreement with previously reported SDP data, which used both neutron and x-ray data. The addition of deuterated and non-deuterated MLV neutron data to the analysis improved lipid backbone information, but did not improve, within experimental uncertainties, the structural information regarding bilayer thickness and area per lipid.

physics.bio-ph

Monolayer Spontaneous Curvature of Raft-Forming Membrane Lipids

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. Spontanous curvatures of both phosphatidylethanolamines and cholesterol were found to be at least a factor of two more negative than those of phosphatidylcholines, whose J0 are closer to zero. Interestingly, a significant positive J0 value (+0.1 1/nm) was retrieved for DPPC at 25 °C. We further determined the temperature dependence of the spontaneous curvatures J0(T) in the range from 15 to 55 \degC, resulting in a quite narrow distribution of -1 to -3 * 10^-3 1/nm°C 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.

cond-mat.soft

Development of a two-dimensional virtual pixel X-ray imaging detector for time-resolved structure research

An interpolating two-dimensional X-ray imaging detector based on a single photon counter with gas amplification by GEM (gas electron multiplier) structures is presented. The detector system can be used for time-resolved structure research down to the microsecond-time domain. The prototype detector has been tested at the SAXS beamline at ELETTRA synchrotron light source with a beam energy of 8 keV to test its capabilities in the rough beamline environment. The imaging performance is examined with apertures and standard diffraction targets. Finally, the application in a time-resolved lipid temperature jump experiment is presented.

physics.ins-det

X-Ray Kinematography of Temperature-Jump Relaxation Probes the Elastic Properties of Fluid Bilayers

The response kinetics of liquid crystalline phosphatidylcholine bilayer stacks to rapid, IR-laser induced temperature jumps has been studied by millisecond time-resolved x-ray diffraction. The system reacts on the fast temperature change by a discrete bilayer compression normal to its surface and a lateral bilayer expansion. Since water cannot diffuse from the excess phase into the interbilayer water region within the 2 ms duration of the laser pulse, the water layer has to follow the bilayer expansion, by an anomalous thinning. Structural analysis of a 20 ms diffraction pattern from the intermediate phase indicates that the bilayer thickness remains within the limits of isothermal equilibrium values. Both, the intermediate structure and its relaxation into the original equilibrium L_(alpha)-phase, depend on the visco-elastic properties of the bilayer/water system. We present an analysis of the relaxation process by an overdamped one-dimensional oscillation model revealing the concepts of Hooke's law for phospholipid bilayers on a supramolecular basis. The results yield a constant bilayer repulsion and viscosity within Hooke's regime suggesting that the hydrocarbon chains act as a buffer for the supplied thermal energy. The bilayer compression is a function of the initial temperature and the temperature amplitude, but is independent of the chain length.

physics.bio-ph

Structural information from multilamellar liposomes at full hydration: full q-range fitting with high quality X-ray data

We present a novel method for analyzing Small Angle X-ray Scattering data on multilamellar phospholipid bilayer systems at full hydration. The method utilizes a modified Caille' 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 new 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.

physics.bio-ph