Search results for "layer"
showing 10 items of 2667 documents
X-ray diffraction of a protein crystal anchored at the air/water interface
1995
We report the first successful in situ x-ray diffraction experiment with a 2D protein array at the lipid/water interface and demonstrate that the order can be controlled via lateral pressure or density. A protein (streptavidin) was bound to a monolayer of biotinylated lipid at the air/water interface, and diffraction of the protein layer could be measured to many orders. Compression of the monolayer changed the diffraction pattern drastically, indicating that the protein structure can be strongly influenced by external parameters like lateral pressure or density. From the width of the peaks, we find that aggregates consisting of as few as 100 monomers contribute to the diffraction. This ind…
Formation of protein multilayers and their competitive replacement based on self-assembled biotinylated phospholipids.
1994
Based on specific recognition processes the build-up of protein multilayers was achieved using streptavidin layers as a docking matrix. For this purpose, streptavidin was organized at biotin-containing monolayers, liposomes, and self-assembled layers on gold. Thus, mixed double and triple layers of streptavidin, Con A, Fab fragments, and hormones were prepared and characterized by fluorescence microscopy and plasmon spectroscopy. Using biotin analogues with lower binding constants several cycles of multilayer formation followed by competitive replacement could be achieved.
Protein-membrane interaction probed by single plasmonic nanoparticles.
2008
We present a nanosized and addressable sensor platform based on membrane coated plasmonic particles and show unequivocally the covering with lipid bilayers as well as the subsequent detection of streptavidin binding to biotinylated lipids. The binding is detected on membrane covered gold nanorods by monitoring the spectral shift by fast single particle spectroscopy (fastSPS) on many particles in parallel. Our approach allows for local analysis of protein interaction with biological membranes as a function of the lateral composition of phase separated membranes.
Streptavidin-coated TiO2 surfaces are biologically inert: Protein adsorption and osteoblast adhesion studies
2011
Non-fouling TiO2 surfaces are attractive for a wide range of applications such as biosensors and medical devices, where biologically inert surfaces are needed. Typically, this is achieved by controlled surface modifications which prevent protein adsorption. For example, polyethylene glycol (PEG) or PEG-derived polymers have been widely applied to render TiO2 surfaces biologically inert. These surfaces have been further modified in order to achieve specific bio-activation. Therefore, there have been efforts to specifically functionalize TiO2 surfaces with polymers with embedded biotin motives, which can be used to couple streptavidin for further functionalization. As an alternative, here a s…
Layer-by-Layer Assembly of a Streptavidin–Fibronectin Multilayer on Biotinylated TiOX
2013
The biomodification of surfaces, especially titanium, is an important issue in current biomedical research. Regarding titanium, it is also important to ensure a specific protein modification of its surface because here protein binding that is too random can be observed. Specific nanoscale architectures can be applied to overcome this problem. As recently shown, streptavidin can be used as a coupling agent to immobilize biotinylated fibronectin (bFn) on a TiO(X) surface. Because of the conformation of adsorbed biotinylated fibronectin on a streptavidin monolayer, it is possible to adsorb more streptavidin and biotinylated fibronectin layers. On this basis, an alternating protein multilayer c…
Surface functionalization and surface recognition: Plasmon optical detection of molecular recognition at self assembled monolayers
1991
The synthesis of biotin- functionalized organic mercaptans and their chemisorption on gold surfaces is described. Biotin bound covalently to self assembled monolayers is recognized by streptavidin from aqueous buffer solutions. Spacer length and packing density of the biotin labels on the organic surface determine the docking kinetics. With a flexible and hydrophilic spacer very fast -diffusion controlled-docking is observed. As an alternative method of self assembly the spreading of organic mercaptans on water surfaces is established. Pressure-area diagrams of different functionalized mercaptans and disulfides are shown and their monolayer properties are discussed.
Molecular mechanisms determining the strength of receptor-mediated intermembrane adhesion
1995
The strength of receptor-mediated cell adhesion is directly controlled by the mechanism of cohesive failure between the cell surface and underlying substrate. Unbinding can occur either at the locus of the specific bond or within the bilayer, which results in tearing the hydrophobic anchors from the membrane interior. In this work, the surface force apparatus has been used to investigate the relationship between the receptor-ligand bond affinities and the dominant mechanism of receptor-coupled membrane detachment. The receptors and ligands used in this study were membrane-bound streptavidin and biotin analogs, respectively, with solution affinities ranging over 10 orders of magnitude. With …
Molecular recognition in biotin-streptavidin systems and analogues at the air-water interface
1992
Abstract Specific interaction between biotin and the protein streptavidin in monolayers of synthetic lipids with biotin headgroups has been shown to lead to formation of highly ordered two-dimensional streptavidin crystals. The same behaviour is observed when using desthiobiotin as lipid headgroup which exhibits a significantly lower binding constant compared with biotin (5 × 10 13 M -1 compared with 10 15 M -1 ). This offers the possibility of detaching competetively the 2D crystalline streptavidin layer by addition of free biotin to the aqueous phase. Use of lipoic acid as lipid headgroup ( K a = 7 × 10 7 M −1 ) leads to formation of small snisotropic protein domains indicating a crystall…
Influence of surface chemistry on the structural organization of monomolecular protein layers adsorbed to functionalized aqueous interfaces.
1993
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…