0000000000113940
AUTHOR
Mathias Lösche
Incorporation of membrane proteins into lipid surface monolayers: Characterization by fluorescence and electron microscopies
The preparation of oriented protein samples is an attractive goal, e.g., to gain more detailed information from spectroscopic experiments. Our approach towards this aim was to prepare monolayers of phospholipids at the air-water interface and to incorporate the proteins into these ordered structures. Subsequently, we used the Langmuir-Boldgett (LB) transfer technique to obtain samples of oriented proteins on solid supports. — Incorporation was achieved by spreading the proteins from a detergent solution onto a prespread lipid monolayer on the water surface. We characterized successful incorporation by in situ fluoresence microscopy and by electron microscopy, and investigated the topology o…
Investigations of the Microstructure of Lipid Interface Films
The phase behaviour of phospholipid monolayers at electrolyte/gas interfaces is studied by fluorescence microscopy. At the LE/LC phase transition, phase separation leads to a Wigner-type lattice structure. The observations are quantified using digital image processing. The results show that the phase transition comprises three different regimes.
Molecular recognition processes at functionalized lipid surfaces: a neutron reflectivity study
The specific binding of proteins to functionalized monolayers on aqueous subphases has been characterized by neutron reflectivity measurements. As a model for the investigation of a recognition process on a molecular length scale, streptavidin (SA) and biotin were chosen because of the high specific affinity between them. Reflectivities from the aqueous (NaCl/H2O or NaCl/D2O) surfaces covered with the biotin-lipid monolayers before and after the adsorption of proteins were collected with a novel, fixed wavelength liquid surface neutron reflectometer. In quantitative terms, binding was found to occur at a biotin surface concentration as low as 1 molecule/1250 A2 (compare to ∼ 1 molecule/40 A…
Structure and elastic properties of smectic liquid crystalline elastomer films
Mechanical measurements, x-ray investigations, and optical microscopy are employed to characterize the interplay of chemical composition, network topology, and elastic response of smectic liquid crystalline elastomers (LCEs) in various mesophases. Macroscopically ordered elastomer films of submicrometer thicknesses were prepared by cross linking freely suspended smectic polymer films. The cross-linked material preserves the mesomorphism and phase transitions of the precursor polymer. The elastic response of the smectic LCE is entropic, and the corresponding elastic moduli are of the order of MPa. In the tilted ferroelectric smectic-C* phase, the network structure plays an important role. Du…
Structural properties of phosphatidylcholine in a monolayer at the air/water interface
Neutron reflectivities of phosphatidylcholine monolayers in the liquid condensed (LC) phase on ultrapure H 2 O and D 2 O subphases have been measured on a Langmuir film balance. Using a dedicated liquid surface reflectometer, reflectivities down to R = 10 -6 in the momentum transfer range Q z = 0–0.4A -1 were accessed. In a new approach, by refining neutron reflectivity data from chain-perdeuterated DPPC-d 62 in combination with x-ray measurements on the same monolayer under similar conditions it is shown that the two techniques mutually complement one another. This analysis leads to a detailed conception of the interface structure. It is found that in the LC phase (which is analogous to th…
Free-standing smectic LC elastomer films
ABSTRACTWe probe the mesophase transitions and layer structures in thin ordered smectic liquid crystalline elastomer films by means of x-ray diffraction and optical microscopy. Oriented elastomer films of submicrometer thickness are produced by crosslinking freely suspended smectic polymer films. After crosslinking, the mesomorphism is similar to that of the precursor polymer. Smectic layers align parallel to the film plane. The layer spacing increases with temperature in the SmC* phase while it decays above the SmC*-SmA transition.
A new liquid surface neutron reflectometer and its application to the study of DPPC in a monolayer at the air/water interface
A constant wavelength neutron reflectometer is described. Using this reflectometer, the neutron reflectivities from phosphatidylcholine monolayers in the highly condensed LC phase on ultra pure H2O and D2O have been measured on a Wilhelmy film balance. The neutron reflectivities have been carefully compared with those obtained by the X-ray method applied to the same monolayer under similar conditions. A new approach to analyzing a combined set of data composed of X-ray and neutron reflectivities has been used. From the analysis it is concluded that despite their limited qz range neutron reflectivities are as essential as X-ray reflectivities for the unique determination of the monolayer str…
Structural Characterization of Molecular Interface Layers Using Neutron and X-Ray Reflectivity Techniques
Synthetic microstructures used in biological research today include a variety of different systems that serve very diverse purposes and require very different fabrication and characterization techniques. One class of microstructures that is at the boundary between life and materials sciences, and certainly has been associated closer to the latter in the past, is that of molecularly structured planar interface films. In fact, Langmuir-Blodgett (LB) films,1 i.e. molecularly layered films comprised of amphiphatic organic molecules on solid substrates, have been used for many years as model systems for the investigation of molecular interactions between chromophores or donor/ acceptor couples, …
Giant lateral electrostriction in ferroelectric liquid-crystalline elastomers
Mechanisms for converting electrical energy into mechanical energy are essential for the design of nanoscale transducers, sensors, actuators, motors, pumps, artificial muscles, and medical microrobots. Nanometre-scale actuation has to date been mainly achieved by using the (linear) piezoelectric effect in certain classes of crystals (for example, quartz), and 'smart' ceramics such as lead zirconate titanate. But the strains achievable in these materials are small--less than 0.1 per cent--so several alternative materials and approaches have been considered. These include grafted polyglutamates (which have a performance comparable to quartz), silicone elastomers (passive material--the constri…
Photoinduced optical anisotropy in organic molecular films controlled by an electric field
Abstract The photoinduced reorientation of dye molecules in molecular films on solid substrates has been controlled with external dc electric fields, leading to solid state structures, which are macroscopically polar, temporally stable at room temperature, and well-defined at the molecular level. A simulation based on the excitation-driven rotational diffusion of the molecules in the potential of their neighbors yielded a realistic model and indicated a collective character of the process. This shows that we are close to a quantitative comprehension of the molecular interactions within these films.
Novel biosensoric devices based on molecular protein hetero-multilayer films
We have developed a novel concept for the modification of technical surfaces with molecularly well-organized layers of bioorganic components. A molecular construction set has been used to implement this concept which is based on molecularly stratified polyelectrolyte films as a structure decoupling protein layers from solid substrates. Utilizing this technology, one can start from a number of different substrates to obtain the same surface structures, on which protein hetero-multilayer films can be prepared to functionalize the interface for (potentially very different) purposes. We have demonstrated the viability of this concept by constructing a biosensor surface that was characterized by…
Photoelectropoling of azobenzene chromophores in molecular films
The photoinduced reorientation and photoelectropoling processes were investigated in molecular films of aliphatically substituted azobenzenes. While polarized illumination leads to a conversion of isotropic samples into anisotropic ones, upon application of external electric fields even macroscopically polar materials are created. The macroscopic polarity can be reversed under suitable experimental conditions. Order parameters as a function of the poling field were determined by Stark spectroscopy. Using atomic force microscopy it is shown that the structure of the sample is well defined on a microscopic length scale, whereas on a macroscopic length scale it is not.
Electro-optical spectroscopy on aggregated chromophore systems in Langmuir-Blodgett films
Linear and quadratic Stark spectra were measured on LB films that contained a hydrophobically substituted cyanine chromophore in binary mixtures with arachidic acid. The external field was applied normal or parallel to the film surface on samples with different internal symmetry. It was found that the orientation of the symmetry axis of the dye molecule is close to the surface normal. A significant number of molecules, 5–10%, are flipped head-to-tail from the main orientation. Parallel to the film surface, large polarizability changes on electronic excitation, Δα ∼ 1000 A3, were observed that reflect electron delocalization over aggregates. At low dye concentrations, large internal electric…
Influence of surface chemistry on the structural organization of monomolecular protein layers adsorbed to functionalized aqueous interfaces.
The molecular organization of streptavidin (SA) bound to aqueous surface monolayers of biotin-functionalized lipids and binary lipid mixtures has been investigated with neutron reflectivity and electron and fluorescence microscopy. The substitution of deuterons (2H) for protons (1H), both in subphase water molecules and in the alkyl chains of the lipid surface monolayer, was utilized to determine the interface structure on the molecular length scale. In all cases studied, the protein forms monomolecular layers underneath the interface with thickness values of approximately 40 A. A systematic dependence of the structural properties of such self-assembled SA monolayers on the surface chemistr…
Optical Studies of Amphiphilic Molecules with Interesting Electro-Optical and Non-Linear Optical Properties
Structural control is a major issue in both life science, investigating the function of the biological machinery, and in materials science, aiming at the design of novel devices. In part one, recent electro-optical investigations of the primary event of photosynthesis on purified protein preparations are described. Part two focuses on structural studies of monolayers at an air/water interface, and of Langmuir-Blodgett multilayers from a new molecule designed for nonlinear optical applications.
Concentration dependence of the electro-optical spectra from two-dimensional aggregated chromophore systems
The linear and the quadratic Stark effects have been employed to characterize aggregate formation of an amphiphilic cyanine dye mixed with arachidic acid in various concentrations within Langmuir-Blodgett multilayers. Two different electrode configurations with the electric field directions parallel and normal to the films have been employed to test the responses of the electronic spectra to external fields. The observations have been used to quantify the induced and static dipole moment changes on excitation of the chromophores. A comparison of the results obtained from different multilayer arrangements is suggestive of inter-layer couplings for the case of head-to-head configurations of t…
Local control of antibody binding to hapten-presenting interfaces: Steric and electrostatic interaction
The binding of labeled antibodies to hapten substituted monolayers at the air/water interface has been studied by means of fluorescence microscopy. Haptens with various spacer lengths between the epitope and a hydrocarbon chain, anchoring the molecule to the interface, have been synthesized. With DMPC,a unspecific binding has been shown to predominate over specific binding due to electrostatic interactions. At high surface pressures the bound antibody is detached because of steric interference with the lipid head groups. Due to a reduction of electrostatic interactions, no unspecific binding is observed to monolayers of cholesterol, which carries a small dipole moment. Mixed monolayers of c…