Incorporation of integrins into artificial planar lipid membranes: characterization by plasmon-enhanced fluorescence spectroscopy

Sinner, Eva‐Kathrin; Reuning, Ute; Kök, Fatma Neşe; Saccá, Barbara; Moröder, Luis; Knoll, Wolfgang; Oesterhelt, Dieter

doi:10.1016/j.ab.2004.05.022

Cited by 41 publications

(50 citation statements)

References 32 publications

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“…In addition, the characterization of solid-supported membranes is feasible by a broad spectrum of biophysical methods, such as electrochemistry, optical spectroscopy and microscopy, and scanning force techniques, to name a few examples. [4][5][6][7][8] One type of solidsupported membrane, peptide-tethered membranes, has been shown to be a suitable platform for functional incorporation of various membrane proteins. [4,9,10] A monomolecular peptide spacer is covalently attached onto a planar metal surface by strong sulfur-gold interactions.…”

mentioning

confidence: 99%

“…The SPFS signal is observed when a fluorophore is close to the surface (150 nm), as reported earlier. [4] The SPFS data presented herein are time-dependent measurements taken at a fixed angle of incidence. An increase in fluorescence over time indicates the interaction of the fluorescently labeled anti-VSV antibody sandwich with the OR5 receptors on the surface, as it is known from the enzyme-linked immunosorbed assay (ELISA).…”

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confidence: 99%

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Incorporation of In Vitro Synthesized GPCR into a Tethered Artificial Lipid Membrane System

et al. 2007

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Controlling the context: A novel strategy for the in vitro expression and immediate post‐translational membrane insertion of complex membrane proteins into an artificial membrane system is described. In this way problems are circumvented that arise when complex membrane proteins are overexpressed, detergent‐solubilized, and subsequently reconstituted into suitable artificial membrane systems for biophysical characterization. GPCR= G‐protein coupled receptor.

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confidence: 99%

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confidence: 99%

Incorporation of In Vitro Synthesized GPCR into a Tethered Artificial Lipid Membrane System

et al. 2007

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“…89 Experiments done with various ECM ligands showed that integrin-ligand binding was specific, which proved that integrin-functionalized membranes are well qualified as sensing devices for the detection of sensible ligand-receptor interactions. 90 A model system that is much closer to cells than planar lipid membranes and SUVs are giant unilamellar vesicles. The only work of integrins reconstitution in GUVs was done by Streicher et al 91 , who had the goal to reconstitute the first steps of integrin-mediated cell spreading (Fig.…”

Section: Reconstitution Of Integrin Into Lipid Membranesmentioning

confidence: 99%

Toward the reconstitution of synthetic cell motility

Siton-Mendelson

Bernheim‐Groswasser

2016

Cell Adhesion & Migration

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Cellular motility is a fundamental process essential for embryonic development, wound healing, immune responses, and tissues development. Cells are mostly moving by crawling on external, or inside, substrates which can differ in their surface composition, geometry, and dimensionality. Cells can adopt different migration phenotypes, e.g., bleb-based and protrusion-based, depending on myosin contractility, surface adhesion, and cell confinement. In the few past decades, research on cell motility has focused on uncovering the major molecular players and their order of events. Despite major progresses, our ability to infer on the collective behavior from the molecular properties remains a major challenge, especially because cell migration integrates numerous chemical and mechanical processes that are coupled via feedbacks that span over large range of time and length scales. For this reason, reconstituted model systems were developed. These systems allow for full control of the molecular constituents and various system parameters, thereby providing insight into their individual roles and functions. In this review we describe the various reconstituted model systems that were developed in the past decades. Because of the multiple steps involved in cell motility and the complexity of the overall process, most of the model systems focus on very specific aspects of the individual steps of cell motility. Here we describe the main advancement in cell motility reconstitution and discuss the main challenges toward the realization of a synthetic motile cell.

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“…The measurements were performed in a home-built apparatus 28 ͑RT-2005, ResTec, Germany͒ equipped with a HeNe laser ͑633 nm͒ and a glass flow cell for liquid handling ͑ϳ20 l volume, Helma, Germany͒. The evaporated gold slide ͑LaSFN9͒ was set on the glass flow cell and mounted on a glass prism ͑LaSFN9͒ in Kretschmann configuration with an index matching oil ͑n = 1.7, Cargille Laboratory., Cleveland, OH͒.…”

Section: H Surface Plasmon Spectroscopymentioning

confidence: 99%

“…The surface coverage of the cBSA was calculated as described 40 ͑⌫ = dc / dn · ⌬n · d͒ and resulted in a surface coverage of 270-280 ng/ cm 2 , using refractive indices of n DPBS = 1.33 and n Protein = 1.4. 28 It should be mentioned that the fitting process considered a uniform film thickness as boundary condition. Figure 2͑d͒ shows the AFM height image of adsorbed cBSA-113 on gold.…”

Section: Gold Surface Coatingmentioning

confidence: 99%

Cationized albumin-biocoatings for the immobilization of lipid vesicles

et al. 2010

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Tethered lipid membranes or immobilized lipid vesicles are frequently used as biomimetic systems. In this article, the authors presented a suitable method for efficient immobilization of lipid vesicles onto a broad range of surfaces, enabling analysis by quantitative methods even under rigid, mechanical conditions-bare surfaces such as hydrophilic glass surfaces as well as hydrophobic polymer slides or metal surfaces such as gold. The immobilization of vesicles was based on the electrostatic interaction of zwitterionic or negatively charged lipid vesicles with two types of cationic chemically modified bovine serum albumin ͑cBSA͒ blood plasma proteins ͑cBSA-113 and cBSA-147͒. Quantitative analysis of protein adsorption was performed as the cBSA coatings were characterized by atomic force microscopy, surface zeta potential measurement, fluorescence microscopy, and surface plasmon spectroscopy, revealing a maximal surface coverage 270-280 ng/ cm 2 for 0.02 mg/ml cBSA on gold. Small unilamellar vesicles as well as giant unilamellar vesicles ͑GUVs͒ were readily immobilized ͑ϳ15 min͒ on cBSA coated surfaces. GUVs with 5-10 mol% negatively charged 1,2,-dipalmitoyl-sn-glycero-3-phosphoglycerol remained stable in liquid for at least 5 weeks.

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Incorporation of integrins into artificial planar lipid membranes: characterization by plasmon-enhanced fluorescence spectroscopy

Cited by 41 publications

References 32 publications

Incorporation of In Vitro Synthesized GPCR into a Tethered Artificial Lipid Membrane System

Incorporation of In Vitro Synthesized GPCR into a Tethered Artificial Lipid Membrane System

Toward the reconstitution of synthetic cell motility

Cationized albumin-biocoatings for the immobilization of lipid vesicles

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