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Biomedical subjects

T Salditt

Publications and source records attributed to T Salditt.

At least 19 recordsLinked to original sources

SARS coronavirus E protein in phospholipid bilayers: an x-ray study.

We investigated the structure of the hydrophobic domain of the severe acute respiratory syndrome E protein in model lipid membranes by x-ray reflectivity and x-ray scattering. In particular, we used x-ray reflectivity to study the location of an iodine-labeled residue within the lipid bilayer. The label imposes spatial constraints on the protein topology. Experimental data taken as a function of protein/lipid ratio P/L and different swelling states support the hairpin conformation of severe acute respiratory syndrome E protein reported previously. Changes in the bilayer thickness and acyl-chain ordering are presented as a function of P/L, and discussed in view of different structural models.

Dimyristoylphosphatidylcholine↗

Electric field unbinding of solid-supported lipid multilayers.

We studied by X-ray reflectivity the behaviour of fully hydrated solid-supported lipid multilayers under the influence of a transverse electric field, under conditions routinely used in the electroformation process. The kinetics of sample loss (unbinding) was measured as a function of the amplitude and frequency of the applied field by monitoring the integrated intensity of the Bragg peaks. We also performed a time-resolved analysis of the intensity of the first Bragg peak and characterized the final state of the sample.

Binding Sites↗

Two-dimensional hard x-ray beam compression by combined focusing and waveguide optics.

A two-dimensionally confining x-ray channel waveguide structure is combined with a high gain Kirkpatrick-Baez prefocusing mirror system yielding a hard x-ray beam with a cross section of 25 x 47 nm(2) (FWHM). Unlike the previously employed resonant beam coupling scheme, the incoming beam is coupled in from the front side of the waveguide and the waveguided beam is no longer accompanied by spurious reflected or transmitted beams. The field distribution in the waveguide channel has been calculated numerically. The calculated transmission and far-field intensity pattern are in good agreement with the experimental results.

Journal Article↗

Collective dynamics of lipid membranes studied by inelastic neutron scattering.

We have studied the collective short wavelength dynamics in deuterated 1,2-dimyristoyl-sn-glycero-3-phoshatidylcholine (DMPC) bilayers by inelastic neutron scattering. The corresponding dispersion relation variant Planck's over 2pi omega(Q) is presented for the gel and the fluid phase of this model system. The temperature dependence of the inelastic excitations indicates a phase coexistence between the two phases over a broad range and leads to a different assignment of excitations from that reported in a preceding inelastic x-ray scattering study [Phys. Rev. Lett. 86, 740 (2001)]]. As a consequence, we find that the minimum in the dispersion relation is actually deeper in the gel than in the fluid phase. Finally, we can clearly identify an additional nondispersive (optical) mode predicted by molecular dynamics simulations [Phys. Rev. Lett. 87, 238101 (2001)]].

Computer Simulation↗

Thermal fluctuations and positional correlations in oriented lipid membranes.

We have determined the positional correlation functions in aligned stacks of fully hydrated phospholipid bilayers from the thermal diffuse scattering measured by nonspecular x-ray reflectivity. While fair agreement can be obtained between experiment and linear smectic theory at length scales above 120 A, significant deviations occur at small r, which are tentatively attributed to collective protrusion modes.

Biomimetic Materials↗

Solid-supported lipid multilayers: structure factor and fluctuations.

We present a theoretical description of the thermal fluctuations in a solid-supported stack of lipid bilayers, for the case of vanishing surface tension gamma = 0 and in the framework of continuous smectic elasticity. The model is successfully used to model the reflectivity profile of a thin (16 bilayers) DMPC sample under applied osmotic pressure and the diffuse scattering from a thick (800 bilayers) stack. We compare our model to previously existing theories.

Computer Simulation↗

Two-dimensional x-ray waveguides and point sources.

We show that resonant coupling of synchrotron beams into suitable nanostructures can be used for the generation of coherent x-ray point sources. A two-dimensionally confining x-ray waveguide structure has been fabricated by e-beam lithography. By shining a parallel undulator beam onto the structure, a discrete set of resonant modes can be excited in the dielectric cavity, depending on the two orthogonal coupling angles between the beam and the waveguide interfaces. The resonant excitation of the modes is evidenced from the characteristic set of coupling angles as well as the observed far-field pattern. The x-ray nanostructure may be used as coherent x-ray point sources with a beam cross section in the nanometer range.

Journal Article↗

Dewetting of solid-supported multilamellar lipid layers.

Thin multilamellar assemblies of neutral lipid bilayers deposited on silicon substrates are shown to be unstable upon hydration. We analyze the stability of these systems taking into account a reduction of the fluctuation-related components of the bilayer interaction potential. The sizes of the patterns observed are consistent with a spinodal dewetting process.

Journal Article↗

Investigation of Structure and Growth of Self-Assembled Polyelectrolyte Layers by X-ray and Neutron Scattering under Grazing Angles.

The structure of self-assembled polyelectrolyte thin films on float glass has been investigated by interface sensitive X-ray and neutron scattering methods. Special emphazis was given to the adsorption process of poly (ethylene imine) and polystyrole sulfonate as an important model system which is often used as a basis for subsequent multilayer buildup. From complementary X-ray and neutron reflectivity data, the vertical film density profile was derived for various growth parameters, including kinetic effects of different adsorption times. In addition to specular reflectivity, we have for the first time employed nonspecular X-ray scattering to study lateral structure parameters in self-assembled polyelectrolyte films. Furthermore, the technique of time-resolved in situ X-ray reflectivity during film growth has been demonstrated and is discussed in view of its future potential. Copyright 2000 Academic Press.

Journal Article↗

Thermal unbinding of highly oriented phospholipid membranes.

We present a temperature dependent x-ray reflectivity study of highly oriented, fully hydrated multilamellar phospholipid membranes. Both the specular and diffuse (nonspecular) x-ray reflectivity were measured for dimyristoyl-sn-glycero-phosphocholine (DMPC) and oleoyl-palmitoyl-sn-glycero-phosphocholine (POPC) on silicon substrates in excess water. In this configuration the repeat distance as well as the fluctuation spectra can be determined as a function of temperature. Both model systems studied exhibit a discontinuous unbinding transition from a substrate bound, multilamellar state to a state of freely dispersed bilayers in water. In the unbound phase a single membrane remains on the substrate.

Dimyristoylphosphatidylcholine↗

Grazing incidence X-ray diffraction of highly aligned phospholipid membranes containing the antimicrobial peptide magainin 2.

We present the first study of grazing incidence X-ray diffraction on a model system of phospholipid membranes and antimicrobial peptides. For this purpose, highly oriented multilamellar samples have been prepared on solid substrates. By this technique, the short-range order of the lipid chains in the fluid L(alpha) phase can be investigated quantitatively, including not only the mean distance between acyl chains, but also the associated correlation length. The short-range order in lecithin is found to be severely affected by the amphiphilic peptide magainin 2.

Amino Acid Sequence↗

Thermal denaturing of bacteriorhodopsin by X-Ray scattering from oriented purple membranes.

We present a temperature-dependent x-ray diffraction study of thin films of purple membranes (PMs) with the native membrane protein bacteriorhodopsin (BR). The high degree of alignment with respect to the silicon substrates allows for the application of modern interface-sensitive scattering techniques. Here we focus on the structural changes of BR in PMs at the thermal denaturing transition. A partial unfolding of the helices is observed rather than the complete unfolding process known from helix to coil transitions. While BR remains threaded into the lipid bilayer in the denatured state, changes in the short-range lateral structures are associated with the partial unfolding of the transmembrane helices.

Bacteriorhodopsins↗

Phase diagram, stability, and overcharging of lamellar cationic lipid-DNA self-assembled complexes.

Cationic lipid-DNA (CL-DNA) complexes comprise a promising new class of synthetic nonviral gene delivery systems. When positively charged, they attach to the anionic cell surface and transfer DNA into the cell cytoplasm. We report a comprehensive x-ray diffraction study of the lamellar CL-DNA self-assemblies as a function of lipid composition and lipid/DNA ratio, aimed at elucidating the interactions determining their structure, charge, and thermodynamic stability. The driving force for the formation of charge-neutral complexes is the release of DNA and lipid counterions. Negatively charged complexes have a higher DNA packing density than isoelectric complexes, whereas positively charged ones have a lower packing density. This indicates that the overcharging of the complex away from its isoelectric point is caused by changes of the bulk structure with absorption of excess DNA or cationic lipid. The degree of overcharging is dependent on the membrane charge density, which is controlled by the ratio of neutral to cationic lipid in the bilayers. Importantly, overcharged complexes are observed to move toward their isoelectric charge-neutral point at higher concentration of salt co-ions, with positively overcharged complexes expelling cationic lipid and negatively overcharged complexes expelling DNA. Our observations should apply universally to the formation and structure of self-assemblies between oppositely charged macromolecules.

Biophysical Phenomena↗

Specular and diffuse scattering of highly aligned phospholipid membranes.

We present a quantitative study of specular and diffuse (non-specular) x-ray and neutron reflectivity from highly aligned phospholipid membranes deposited on solid substrates. The height-height correlation function could be obtained from the diffuse scattering without further model assumptions. The results differ significantly from the linear theory of smectic elasticity. We argue that the diffuse scattering is dominated by static liquid-crystalline defects, rather than thermal fluctuations.

1,2-Dipalmitoylphosphatidylcholine↗

An inverted hexagonal phase of cationic liposome-DNA complexes related to DNA release and delivery.

A two-dimensional columnar phase in mixtures of DNA complexed with cationic liposomes has been found in the lipid composition regime known to be significantly more efficient at transfecting mammalian cells in culture compared to the lamellar (LalphaC) structure of cationic liposome-DNA complexes. The structure, derived from synchrotron x-ray diffraction, consists of DNA coated by cationic lipid monolayers and arranged on a two-dimensional hexagonal lattice (HIIC). Two membrane-altering pathways induce the LalphaC --> HIIC transition: one where the spontaneous curvature of the lipid monolayer is driven negative, and another where the membrane bending rigidity is lowered with a new class of helper-lipids. Optical microscopy revealed that the LalphaC complexes bind stably to anionic vesicles (models of cellular membranes), whereas the more transfectant HIIC complexes are unstable and rapidly fuse and release DNA upon adhering to anionic vesicles.

Birefringence↗

Structure of DNA-cationic liposome complexes: DNA intercalation in multilamellar membranes in distinct interhelical packing regimes.

Cationic liposomes complexed with DNA (CL-DNA) are promising synthetically based nonviral carriers of DNA vectors for gene therapy. The solution structure of CL-DNA complexes was probed on length scales from subnanometer to micrometer by synchrotron x-ray diffraction and optical microscopy. The addition of either linear lambda-phage or plasmid DNA to CLs resulted in an unexpected topological transition from liposomes to optically birefringent liquid-crystalline condensed globules. X-ray diffraction of the globules revealed a novel multilamellar structure with alternating lipid bilayer and DNA monolayers. The lambda-DNA chains form a one-dimensional lattice with distinct interhelical packing regimes. Remarkably, in the isoelectric point regime, the lambda-DNA interaxial spacing expands between 24.5 and 57.1 angstroms upon lipid dilution and is indicative of a long-range electrostatic-induced repulsion that is possibly enhanced by chain undulations.

Bacteriophage lambda↗