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Monika Majerowicz

Publications and source records attributed to Monika Majerowicz.

3 recordsLinked to original sources

Membrane binding of a lipidated N-Ras protein studied in lipid monolayers.

The adsorption of doubly lipidated full-length N-Ras protein on 1,2-dipalmitoyl-sn-phosphatidylcholine (DPPC) monolayers was studied by lateral pressure analysis, grazing incidence X-ray diffraction (GIXD), and specular reflectivity (XR). N-Ras protein adsorbs to the DPPC monolayer (lateral pressure of 20 mN/m) from the subphase thereby increasing the lateral pressure in the monolayer by 4 mN/m. The protein insertion does not alter the tilt angle and structure of the lipid molecules at the air/water interface but influences the electron density profile of the monolayer. Further, electron density differences into the subphase were observed. The Fresnel normalized reflectivity could be reconstructed in the analysis using box models yielding electron density profiles of the DPPC monolayer in the absence and in the presence of N-Ras protein. The electron density profiles of the DPPC monolayer in the presence of Ras showed clear intensity variations in the headgroup/glycerol/upper chain region, the so-called interface region where previous bilayer studies had confirmed Ras binding.

1,2-Dipalmitoylphosphatidylcholine↗

Phospholipid mesophases at solid interfaces: in-situ X-ray diffraction and spin-label studies.

In this work, we report on recent investigations, both on the global and on the local molecular architecture of supported phospholipid model membranes. A brief theoretical introduction explains how global structural information on supramolecular lipid ensembles can be retrieved from surface X-ray diffraction measurements as well as how spin-label electron paramagnetic resonance spectroscopy (EPR) provides complementary information on the local environment of probe molecules. The combination of especially designed X-ray cells with the technique of small- and wide-angle X-ray surface scattering makes it possible to explore various fields of lipid research and its applications. Examples for different physico-chemical conditions are presented: (i) in situ chemistry under excess of water conditions demonstrating how solid-supported lipid films sense salinity, (ii) the 3D electron density reconstruction of a vesicle-fusion intermediate under controlled humidity, and (iii) complementary temperature and pressure effects on oriented phospholipid samples. Further, special attention has been given to the influence of different film preparation techniques with respect to quality and the defect structure manifestation. To resolve the proportions and local properties of defects in a hydrated lipid-deposited surface, spin-label EPR was applied. The results from 9.6 GHz EPR as well as from 1.2 GHz EPR suggest the alignment to be in the range between 30% and 80%. In addition, slow time-dependent EPR measurements point to nano-structural rearrangements due to water flow and reduction of alignment quality.

Cell Membrane↗

In situ sensing of salinity in oriented lipid multilayers by surface X-ray scattering.

The influence of LiCl solutions on liposomal and surface-supported phosphatidylcholine/water systems (dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), respectively) has been studied by small-angle X-ray techniques. In liposomal dispersions of DPPC, an osmotically stressed liquid-crystalline phase, denoted as Lalpha osm, forms readily after rapid mixing with salt solutions. The transition from Lalpha -->Lalpha osm proceeds in two steps. The first step takes place within seconds and is due to water diffusion from the liposome into the bulk solution. The second, slower process (minutes) can be attributed to the relaxation of initially deformed intermediate liposomes into spherical ones. In experiments with aligned lipid bilayers supported on silicon wafers, it was possible to reproducibly exchange different concentrations of LiCl solutions on a single sample and to determine the lattice changes by time-resolved X-ray scattering at grazing incidence. Independently of the deposition technique (spray- or spin-coating, respectively), none of the investigated POPC samples displayed an osmotically stressed liquid-crystalline phase. While liposomes can be considered nearly defect-free, supported bilayer stacks show a high abundance of defects, such as oily streaks typical of the Lalpha phase. Thus, the alkali ions are free to diffuse into the interbilayer water regions and to cause a slight increase of the bilayer separation (about 1 Angstroms). It is concluded that low to medium concentrations of Li+ ions partially screen the attractive van der Waals force between adjacent membrane layers. However, upon annealing the defect regions or regions of high curvature in the oriented lipid matrix, e.g. by low amounts of oleyl alcohol (OA), the system is able to sense osmotic stress upon addition of a salt solution.

1,2-Dipalmitoylphosphatidylcholine↗