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Biomedical subjects

Andreas Janshoff

Publications and source records attributed to Andreas Janshoff.

At least 19 recordsLinked to original sources
PubMed2006

Electrically induced deformation of giant liposomes monitored by thickness shear mode resonators.

Thickness shear mode resonators are capable of registering small changes in the thickness and viscoelastic properties of ultrathin films attached to their surface. It was found that it is possible to monitor the deformation of surface-bound giant liposomes by applying an electric field with small amplitudes. Changes in the apparent height of attached vesicles in the nanometer range were easily detected as a function of lipid composition. Increasing the bending modulus by adding cholesterol results in a significantly reduced deformation from 16.8 nm (5% cholesterol) down to 3.2 nm (20% cholesterol), rendering this new method a robust and sensitive tool to detect the bending elasticity of liposomes on small length scales. Deformation could be further suppressed by adding anchor groups (biotinylated lipids), resulting in a strongly flattened liposome on an avidin-coated resonator.

Cholesterol↗
PubMed2006

Control of CaCO3 crystallization by demixing of monolayers.

In this paper we describe how to template a demixed monolayer into a spatially patterned inorganic replica. For this purpose a new amphiphilic monomer was synthesized which can be polymerized both in solution and in the monolayer of a Langmuir-Blodgett (LB) trough. Since it inhibits the crystallization of CaCO3, it can be used--in combination with stearic acid (nucleation-promotor)--to control CaCO3 crystals formed under the monolayer. Investigations of the two-component monolayer (Langmuir isotherms and AFM measurements of transferred films) show--in the biphasic region--demixing in solid analogue stearic acid domains and the liquid analogue phase of the monomer. Crystallization of CaCO3 starts under the stearic acid domains whose size varies from less than 100 nm to several tens of micrometers. The addition of poly(acrylic acid) into the subphase hinders the three-dimensional growth of CaCO3 crystals from the monolayer into the solution. Thus, it becomes possible to transfer the pattern of the demixed domains into an inorganic replica of CaCO3.

Acrylic Resins↗
PubMed2006

Cooperative adsorption of ezrin on PIP2-containing membranes.

By means of the quartz crystal microbalance (QCM) and scanning force microscopy (SFM), the adsorption of ezrin, a member of the ezrin/radixin/moesin protein family, on l-alpha-phosphatidylinositol-4,5-bisphosphate (PIP(2)) containing solid-supported membranes was investigated. An increase in the PIP(2) content in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes resulted in an increased amount of bound ezrin strongly supporting the crucial role of PIP(2) for ezrin recruitment to membranes. No ezrin adsorption to membranes composed of pure POPC was detected. To characterize the binding process in more detail, the kinetics and reversibility of ezrin adsorption were investigated by the QCM technique, showing that the protein remains partly bound after rinsing with pure buffer, which we suspected to be a result of lateral interactions between the proteins. SFM images revealed the formation of two-dimensional ezrin clusters on PIP(2)-doped POPC membranes. Time-elapsed SFM images show that the growth of protein domains occurs from a few nucleation sites. The QCM data in conjunction with the results obtained by SFM led us to propose that the binding process of ezrin occurs in a positive cooperative manner. When lateral interactions of the proteins on the membrane were taken into account, we were able to simulate the kinetics obtained from time-resolved QCM readouts by employing a model developed by Minton. On the basis of the kinetic analysis, we were also able to reconstruct the adsorption isotherm.

Adsorption↗
PubMed2006

Cell motility probed by noise analysis of thickness shear mode resonators.

The quartz crystal microbalance (QCM) technique is an emerging bioanalytical tool to study the behavior of animal cells in vitro. Due to the high interfacial sensitivity of thickness shear mode (TSM) resonators it is possible to monitor the formation and breakage of cell-matrix interactions and changes in viscoelasticity of the cell bodies, as well as minute cell volume alterations by the time course of their resonance frequency even with millisecond time resolution. We found that mammalian MDCK-II cells grown on TSM resonators impose characteristic fluctuations on the resonance frequency, which are a quantitative indicator for dynamic activities of the cells on the surface and report on their vitality and motility. Applying noise analysis to the fluctuating resonance frequency allows one to quantify the response of the cells to environmental changes such as osmotic stress, addition of fixation reagents, or the influence of drugs such as cytochalasin D. The corresponding power density spectra of the noise imposed on the resonance frequency by the dynamic activities of the cells show a characteristic resonance at 1-2 Hz, which can be substantially altered by osmotic stress, fixation agents, or cytochalasin D. Comparison of QCM-based fluctuation readings with electric cell--substrate impedance sensing (ECIS)--a well-established technique to monitor cell dynamics-provides substantially different results, indicating that both techniques may complement each other with respect to their biological information. Whereas ECIS readings report solely on cell shape changes, QCM-based fluctuation analysis is also influenced by fluctuations in the viscoelasticity of the cell bodies.

Animals↗
PubMed2006

Facile synthesis and characterization of functionalized, monocrystalline rutile TiO2 nanorods.

Functionalized, monocrystalline rutile TiO2 nanorods were prepared from TiCl4 in aqueous solution under acidic conditions in the presence of dopamine, followed by aging and hydrothermal treatment at 150 degrees C. The surface-bound organic ligand controls the morphology as well as the crystallinity and the phase selection of TiO2. The presence of monocrystalline rutile TiO2 was confirmed by X-ray powder diffraction and HRTEM investigations. The as-prepared nanorods are soluble in water at pH <3. The surface functionalization was analyzed by IR and 1H NMR, confirming the presence of dopamine on the surface. The surface amine groups can be tailored further with functional molecules such as dyes. Confocal laser scanning microscopy (CLSM) was used to characterize the binding of the fluorescent dye 4-chloro-7-nitrobenzofurazan (NBD) to the functionalized surface of the TiO2 nanorods.

Journal Article↗
PubMed2006

Mechanical properties of pore-spanning lipid bilayers probed by atomic force microscopy.

We measure the elastic response of a free-standing lipid membrane to a local indentation by using an atomic force microscope. Starting point is a planar gold-coated alumina substrate with a chemisorbed 3-mercaptopropionic acid monolayer displaying circular pores of very well defined and tunable size, over which bilayers composed of N,N,-dimethyl-N,N,-dioctadecylammonium bromide or 1,2-dioleoyl-3-trimethylammonium-propane chloride were spread. Centrally indenting these "nanodrums" with an atomic force microscope tip yields force-indentation curves, which we quantitatively analyze by solving the corresponding shape equations of continuum curvature elasticity. Since the measured response depends in a known way on the system geometry (pore size, tip radius) and on material parameters (bending modulus, lateral tension), this opens the possibility to monitor local elastic properties of lipid membranes in a well-controlled setting.

Computer Simulation↗
PubMed2006

Transport across artificial membranes-an analytical perspective.

Biosensors that make use of transport processes across lipid membranes are very rare even though a stimulus, the binding of a single analyte molecule, can enhance the sensor response manifold if the analyte leads to the transport of more than one ion or molecule across the membrane. Prerequisite for a proper function of such membrane based biosensors is the formation of lipid bilayers attached to a support that allow for the insertion of membrane peptides and proteins in a functional manner. In this review, the current state of the art technologies to obtain lipid membranes on various supports are described. Solid supported membranes on transparent and electrically conducting surfaces, lipid bilayers on micromachined apertures and on porous materials are discussed. The focus lies on the applicability of such membranes for the investigation of transport phenomena across lipid bilayers facilitated by membrane embedded peptides, channel proteins and transporters. Carriers and channel forming peptides, which are easy to handle and rather robust, are used frequently to build up membrane based biosensors. However, channel forming proteins and transporters are more difficult to insert functionally and thus, there are yet only few examples that demonstrate the applicability of such systems as biosensor devices.

Animals↗
PubMed2006

Adsorption and fluctuations of giant liposomes studied by electrochemical impedance measurements.

The present study describes a novel approach based on electrochemical impedance measurements to follow the adsorption of giant liposomes on protein-coated solid surfaces with a time resolution in the order of seconds. The technical key features are circular gold-film electrodes as small as a few hundred micrometers in diameter and measurements of the electrode capacitance using AC signals in the kilohertz regime. Using Monte Carlo simulations, we were able to support the experiments and extract the rate constant of liposome adsorption. Besides monitoring the adsorption of liposomes on protein-coated surfaces, we also applied this technique to study shape fluctuations of the adsorbed vesicles and compared the corresponding power spectra with those recorded for hard particles and living animal cells.

Adsorption↗
PubMed2006

Partially reversible adsorption of annexin A1 on POPC/POPS bilayers investigated by QCM measurements, SFM, and DMC simulations.

The kinetics of annexin A1 binding to solid-supported lipid bilayers consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS; 4:1) has been investigated as a function of the calcium ion concentration in the bulk phase. Quartz crystal microbalance measurements in conjunction with scanning force microscopy, fluorescence microscopy, and computer simulations indicate that at a given Ca2+ concentration annexin A1 adsorbs irreversibly on membrane domains enriched in POPS. By contrast, annexin A1 adsorbs reversibly on the POPC-enriched phase, which is composed of single POPS molecules embedded within a POPC matrix. The overall area occupied by the POPS-enriched phase is controlled by the CaCl2 concentration. Monte Carlo simulations suggest that the area of the POPS-enriched phase increases by a factor of 7 when the Ca2+ concentration is changed from 0.01 to 1 mM.

Adsorption↗
PubMed2005

Microstructuring of phospholipid bilayers on gold surfaces by micromolding in capillaries.

Microstructuring of lipid bilayers on gold surfaces was achieved by micromolding in capillaries employing chemically modified polydimethylsiloxane (PDMS). Microfluidic networks of PDMS were prepared by micromolding and functionalized with thiol end-groups using 3-mercaptopropyltrimethoxysilane. The PDMS stamps were firmly attached to the gold substrate via quasi-covalent linkage providing a tight seal, a prerequisite for establishing individual addressable capillaries. Bilayers composed of POPC/POPG were subsequently prepared on microstructured self assembly monolayers of 11-amino-1-undecanethiol via strong electrostatic interactions. This way it is possible to generate individually addressable lipid bilayers on gold surfaces, a procedure, which is of widespread interest for investigating protein lipid interactions with microscopic techniques.

Dimethylpolysiloxanes↗
PubMed2005

Label-free detection of protein-ligand interactions by the quartz crystal microbalance.

In recent years the quartz crystal microbalance (QCM) has been accepted as a powerful technique to monitor adsorption processes at interfaces in different chemical and biological research areas. In the last decade, the investigation of adsorption of biomolecules on functionalized surfaces turned out to be one of the paramount applications of the QCM comprising the interaction of nucleic acids, specific molecular recognition of protein-receptor couples, and antigen-antibody reactions realized in immunosensors. The advantage of the QCM technique is that it allows for a label free detection of molecules. This is a result of the fact that the frequency response of the quartz resonator is proportional to the increase in thickness of the adsorbed layer. However, in recent years it became more and more evident that quartz resonators used in fluids are more than mere mass or thickness sensors. The sensor response is also influenced by viscoelastic properties of the adhered biomaterial, surface charges of adsorbed molecules and surface roughness. These phenomena have been used to get new insights in the adhesion process of living cells and to understand their response to pharmacological substances by determining morphological changes of the cells. In this chapter we describe a protocol to explore the kinetics and thermodynamics of specific interactions of different proteins such as lectins and annexins with their ligands using receptor bearing solid supported lipid bilayers.

Animals↗
PubMed2004

Monitoring the formation of biosilica catalysed by histidine-tagged silicatein.

Surface bound silicatein retains its biocatalytic activity, which was demonstrated by monitoring the immobilisation of silicatein using a histidine-tag chelating anchor and the subsequent biosilicification of SiO(2) on surfaces by surface plasmon resonance spectroscopy, atomic force microscopy and scanning electron microscopy.

Catalysis↗