Fluctuations actives et interactions électrostatiques dans les membranes lipidiques flottantes

The main project of this work was focused on the investigation of out-of-equilibrium fluctuation of phospholipid membranes induced by light-activated transmembrane protein bacteriorhodopsin (BR). A robust protocol for the BR incorporation into the membrane-mimic systems was developed and the induced structural changes caused by BR incorporation and activation with light were probed by means of neutron and X-ray specular and off-specular reflectometry. The reversible effect of light illumination on the protein activity (on /off) via its effect on the bilayer structure and fluctuation spectrum was demonstrated. These results open the way to investigate the active fluctuation spectrum of a planar membrane-protein system and to access the physical properties of the active membrane. The aim of the second project was to investigate the interaction between highly negatively charged DPPS lipid bilayers. We fully characterized the structure of the system and clearly demonstrated that attractive interactions existed between charged bilayers, in good agreement with Strong-Coupling theory.Le projet principal de ce travail a porté sur l'étude des fluctuations hors d'équilibres de membranes lipidiques induites par la bactériorhodopsine (BR), une protéine transmembranaire activée par la lumière. Un protocole robuste pour l'incorporation de BR dans les systèmes mimétiques de membrane a été développé et les changements structurels induits par l'incorporation et l'activation de la BR ont été étudiés en détail par réflectivité spéculaire (neutrons et rayons X) et par réflectivité hors-spéculaire (rayons X). Nous avons pu observer un effet réversible induit par l'activité de la protéine sur la structure de la membrane et sur ses fluctuations. Ces résultats ouvrent la voie à l’étude du spectre des fluctuations hors-équilibres d’un système protéo-membranaire planaire, et à l’accès aux propriétés physiques de la membrane active. Dans un second projet nous avons étudié l’interaction entre deux bicouches lipidiques fortement chargées. Nous avons finement caractérisé la structure du système et clairement démontré que des interactions attractives existaient entre les bicouches chargées, en accord avec la théorie de couplage fort

Phase Behavior and Miscibility in Two-Component Glycolipid Monolayers

Glycolipids are known to be involved in the formation of ordered functional domains in biological membranes. Since the structural characterization of such domains is difficult, most studies have so far dealt with lipid mixtures containing only one glycolipid component at a time, although biological membranes usually contain several glycolipid species, which can result in more complex structures and phase behavior. Here, we combine classical isotherm measurements with surface-sensitive grazing-incidence X-ray diffraction to investigate the phase behavior and miscibility in Langmuir monolayers of binary glycolipid mixtures. We find that the phase behavior has a subtle dependence on the saccharide headgroup chemistry. For compatible chemistries, molecular superlattice structures formed by one of the glycolipid species are conserved and can host foreign glycolipids up to a defined stoichiometry. In contrast, for sterically incompatible saccharide chemistries, the superlattice is lost even if both species are able to form such structures in their pure forms. Our results suggest that related phenomena may play important roles also in biological contexts

The Complete Phase Diagram of Monolayers of Enantiomeric N-Stearoyl-threonine Mixtures with Preferred Heterochiral Interactions

Langmuir monolayers of chiral amphiphiles are well-controlled model systems for the investigation of phenomena related to stereochemistry. Here, we have investigated mixed monolayers of one pair of enantiomers (l and d) of the amino-acid-based amphiphile N-stearoyl-threonine. The monolayer characteristics were studied by pressure–area isotherm measurements and grazing incidence X-ray diffraction (GIXD) over a wide range of mixing ratios defined by the d-enantiomer mole fraction xD. While the isotherms provide insights into thermodynamical aspects, such as transition pressure, compression/decompression hysteresis, and preferential homo- and heterochiral interactions, GIXD reveals the molecular structural arrangements on the Ångström scale. Dominant heterochiral interactions in the racemic mixture lead to compound formation and the appearance of a nonchiral rectangular lattice, although the pure enantiomers form a chiral oblique lattice. Miscibility was found to be limited to mixtures with 0.27 ≲ xD ≲ 0.73, as well as to both outer edges (xD ≲ 0.08 and xD ≳ 0.92). Beyond this range, coexistence of oblique and rectangular lattices occurs, as is clearly seen in the GIXD patterns. Based on the results, a complete phase diagram with two eutectic points at xD ≈ 0.25 and xD ≈ 0.75 is proposed. Moreover, N-stearoyl-threonine was found to have a strong tendency to form a hydrogen-bonding network between the headgroups, which promotes superlattice formation

The Complete Phase Diagram of Monolayers of Enantiomeric N-Stearoyl-threonine Mixtures with Preferred Heterochiral Interactions

Langmuir monolayers of chiral amphiphiles are well-controlled model systems for the investigation of phenomena related to stereochemistry. Here, we have investigated mixed monolayers of one pair of enantiomers (L and D) of the amino-acidbased amphiphile N-stearoyl-threonine. The monolayer characteristics were studied by pressure−area isotherm measurements and grazing incidence X-ray diffraction (GIXD) over a wide range of mixing ratios defined by the D-enantiomer mole fraction $x_D$. While the isotherms provide insights into thermodynamical aspects, such as transition pressure, compression/decompression hysteresis, and preferential homo- and heterochiral interactions, GIXD reveals the molecular structural arrangements on the Ångström scale. Dominant heterochiral interactions in the racemic mixture lead to compound formation and the appearance of a nonchiral rectangular lattice, although the pure enantiomers form a chiral oblique lattice. Miscibility was found to be limited to mixtures with 0.27 ≲ $x_D$ ≲ 0.73, as well as to both outer edges ($x_D$ ≲ 0.08 and $x_D$ ≳ 0.92). Beyond this range, coexistence of oblique and rectangular lattices occurs, as is clearly seen in the GIXD patterns. Based on the results, a complete phase diagram with two eutectic points at $x_D$ ≈ 0.25 and $x_D$ ≈ 0.75 is proposed. Moreover, N-stearoyl-threonine was found to have a strong tendency to form a hydrogen-bonding network between the headgroups, which promotes superlattice formation

Метод оцінювання інформативності еталонних даних геофізичного поля

This article presents an integral index for informativity estimation of geophysical field template. Informativity estimation algorithm and software for its realization are developed. The experiments on a series of pictures of different regions (field, forest, city, etc.) confirmed the effectiveness and accuracy of the given algorithm.Запропоновано інтегральний параметр для оцінювання інформативності еталонних даних геофізичного поля. Розроблено алгоритм та програмне забезпечення для його реалізації. Експерименти на наборі зображень різних місцевостей (поле, ліс, місто) підтвердили ефективність та точність даного алгоритму

Метод оцінювання інформативності еталонних даних геофізичного поля

This article presents an integral index for informativity estimation of geophysical field template. Informativity estimation algorithm and software for its realization are developed. The experiments on a series of pictures of different regions (field, forest, city, etc.) confirmed the effectiveness and accuracy of the given algorithm.Запропоновано інтегральний параметр для оцінювання інформативності еталонних даних геофізичного поля. Розроблено алгоритм та програмне забезпечення для його реалізації. Експерименти на наборі зображень різних місцевостей (поле, ліс, місто) підтвердили ефективність та точність даного алгоритму

pH-Dependent physicochemical properties of ornithine lipid in mono- and bilayers

In certain bacteria, phosphatidylethanolamine lipids (PEL) get largely replaced by phosphate-free ornithine lipids (OL) under conditions of phosphate starvation. It has so far been unknown how much these two lipid types deviate in their physicochemical properties, and how strongly bacteria thus have to adapt in order to compensate for the difference. Here, we use differential scanning calorimetry, X-ray scattering, and X-ray fluorescence to investigate the properties of OL with saturated C14 alkyl chains in mono- and bilayers. OL is found to have a greater tendency than chain-analogous PEL to form ordered structures and, in contrast to PEL, even a molecular superlattice based on a hydrogen bonding network between the headgroups. This superlattice is virtually electrically uncharged and persists over a wide pH range. Our results indicate that OL and PEL behave very differently in ordered single-component membranes but may behave more similarly in fluid multicomponent membranes

Characterization of Lipid Bilayers Adsorbed to Functionalized Air/Water Interfaces

Lipid bilayers immobilized in planar geometries, such as solid-supported or "floating" bilayers, have enabled detailed studies of biological membranes with numerous experimental techniques, notably x-ray and neutron reflectometry. However, the presence of a solid support also has disadvantages as it complicates the use of spectroscopic techniques as well as surface rheological measurements that would require surface deformations. Here, in order to overcome these limitations, we investigate lipid bilayers adsorbed to inherently soft and experimentally well accessible air/water interfaces that are functionalized with Langmuir monolayers of amphiphiles. The bilayers are characterized with ellipsometry, X-ray scattering, and X-ray fluorescence. Grazing-incidence X-ray diffraction reveals that lipid bilayers in a chain-ordered state can have significantly different structural features than regular Langmuir monolayers of the same composition. Our results suggest that bilayers at air/water interfaces may be well suited for fundamental studies in the field of membrane biophysics

Characterization of lipid bilayers adsorbed to functionalized air/water interfaces

Lipid bilayers immobilized in planar geometries, such as solid-supported or “floating” bilayers, have enabled detailed studies of biological membranes with numerous experimental techniques, notably X-ray and neutron reflectometry. However, the presence of a solid support also has disadvantages as it complicates the use of spectroscopic techniques as well as surface rheological measurements that would require surface deformations. Here, in order to overcome these limitations, we investigate lipid bilayers adsorbed to inherently soft and experimentally well accessible air/water interfaces that are functionalized with Langmuir monolayers of amphiphiles. The bilayers are characterized with ellipsometry, X-ray scattering, and X-ray fluorescence. Grazing-incidence X-ray diffraction reveals that lipid bilayers in a chain-ordered state can have significantly different structural features than regular Langmuir monolayers of the same composition. Our results suggest that bilayers at air/water interfaces may be well suited for fundamental studies in the field of membrane biophysics

Attractive Interaction between Fully Charged Lipid Bilayers in a Strongly Confined Geometry

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