Solid-Supported Biomolecules on Modified Silica Surfaces—A Tool for Fast Physicochemical Characterization and High-Throughput Screening

Loidl-Stahlhofen, Angelika; Schmitt, Johannes; Nöller, Joachim; Hartmann, Thorsten; Brodowsky, Hanna M.; Schmitt, Walter; Keldenich, Jörg

doi:10.1002/1521-4095(200112)13:23<1829::aid-adma1829>3.0.co;2-3

Cited by 60 publications

(12 citation statements)

References 30 publications

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“…2 A typical application of these supported lipid membranes is to serve as a scaffold for membrane-associated biological functions; [3][4][5][6][7][8][9][10] for example, in our previous work we have used supported lipid layers as a matrix for the attachment of viral functions to colloidal particles via membrane fusion. 11 The importance of supported membranes from both the scientific and technological point of view has led to extensive research in the past.…”

Section: Introductionmentioning

confidence: 99%

Lipid layers on polyelectrolyte multilayer supports

et al. 2008

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The mechanism of formation of supported lipid layers from phosphatidylcholine and phosphatidylserine vesicles in solution on polyelectrolyte multilayers was studied by a variety of experimental techniques. The interaction of zwitterionic and acidic lipid vesicles, as well as their mixtures, with polyelectrolyte supports was followed in real time by micro-gravimetry. The fabricated lipid-polyelectrolyte composite structures on top of multilayer coated colloidal particles were characterized by flow cytometry and imaging techniques. Lipid diffusion over the macroscopic scale was quantified by fluorescence recovery after photobleaching, and the diffusion was related to layer connectivity. The phospholipid-polyelectrolyte binding mechanism was investigated by infrared spectroscopy. A strong interaction of polyelectrolyte primary amino groups with phosphate and carboxyl groups of the phospholipids, leading to dehydration, was observed. Long-range electrostatic attraction was proven to be essential for vesicle spreading and rupture. Fusion of lipid patches into a homogeneous bilayer required lateral mobility of the lipids on the polyelectrolyte support. The binding of amino groups to the phosphate group of the zwitterionic lipids was too weak to induce vesicle spreading, but sufficient for strong adsorption. Only the mixture of phosphatidylcholine and phosphatidylserine resulted in the spontaneous formation of bilayers on polyelectrolyte multilayers. The adsorption of phospholipids onto multilayers displaying quarternary ammonium polymers produced a novel 3D lipid polyelectrolyte structure on colloidal particles.

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Section: Introductionmentioning

confidence: 99%

Lipid layers on polyelectrolyte multilayer supports

et al. 2008

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“…14 In another report, similar bilayer-coated beads were used to study phosphoinositide-specific phospholipase activity using laser trap-based microelectrophoresis. 15 Expansion of these capabilities through high throughput parallel and multiplex strategies offers great possibilities for biosensor technology; 16,17 however, functional reconstitution of complex transmembrane proteins into this platform has yet to be demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Nanoporous Microbead Supported Bilayers: Stability, Physical Characterization, and Incorporation of Functional Transmembrane Proteins

et al. 2007

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The introduction of functional transmembrane proteins into supported bilayer-based biomimetic systems presents a significant challenge for biophysics. Among the various methods for producing supported bilayers, liposomal fusion offers a versatile method for the introduction of membrane proteins into supported bilayers on a variety of substrates. In this study, the properties of protein containing unilamellar phosphocholine lipid bilayers on nanoporous silica microspheres are investigated. The effects of the silica substrate, pore structure, and the substrate curvature on the stability of the membrane and the functionality of the membrane protein are determined. Supported bilayers on porous silica microspheres show a significant increase in surface area on surfaces with structures in excess of 10 nm as well as an overall decrease in stability resulting from increasing pore size and curvature.Comparison of the liposomal and detergent-mediated introduction of purified bacteriorhodopsin (bR) and the human type 3 serotonin receptor (5HT3R) are investigated focusing on the resulting protein function, diffusion, orientation, and incorporation efficiency. In both cases, functional proteins are observed; however, the reconstitution efficiency and orientation selectivity are significantly enhanced through detergent-mediated protein reconstitution. The results of these experiments provide a basis for bulk ionic and 4 fluorescent dye-based compartmentalization assays as well as single-molecule optical and single-channel electrochemical interrogation of transmembrane proteins in a biomimetic platform. 5

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“…Biacore has produced a variant of their surface plasmon resonance (SPR) technology where a dextran coating on their SPR electrode is modified with surface lipids that promote the fusion of solubilized GPCRs [20]. Nimbus Inc. reconstitutes transmembrane proteins into lipid bilayers supported on porous silica cores [21], while Biovectors Therapeutics S.A. uses polysaccharide cores [22]. Our method represents an alternative approach to developing binding assays in which GPCRs are reconstituted into an easily manipulated lipid bilayer environment.…”

Section: Resultsmentioning

confidence: 99%

Characterization of radioligand binding to a transmembrane receptor reconstituted into Lipobeads

Park

Buck

et al. 2004

FEBS Letters

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Lipobeads are hydrogel beads surrounded by a lipid bilayer membrane and have been developed to act as a cell analogue. The FLAG-tagged M 2 muscarinic receptor was incorporated onto the surface of the Lipobead by incubating pre-Lipobeads with proteoliposomes containing the receptor. Receptors reconstituted onto the surface of the Lipobeads were functional in that they bound the antagonists quinuclidinylbenzilate and scopolamine with characteristic muscarinic affinities. This demonstrates the feasibility of using Lipobeads to study the binding properties of the M 2 muscarinic receptor and offers a promising approach to the study of transmembrane protein biology in general.

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Solid-Supported Biomolecules on Modified Silica Surfaces—A Tool for Fast Physicochemical Characterization and High-Throughput Screening

Cited by 60 publications

References 30 publications

Lipid layers on polyelectrolyte multilayer supports

Lipid layers on polyelectrolyte multilayer supports

Nanoporous Microbead Supported Bilayers: Stability, Physical Characterization, and Incorporation of Functional Transmembrane Proteins

Characterization of radioligand binding to a transmembrane receptor reconstituted into Lipobeads

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