DC Field | Value | Language |
dc.contributor.author | Roder, Friedrich | |
dc.contributor.author | Waichman, Sharon | |
dc.contributor.author | Paterok, Dirk | |
dc.contributor.author | Schubert, Robin | |
dc.contributor.author | Richter, Christian | |
dc.contributor.author | Liedberg, Bo | |
dc.contributor.author | Piehler, Jacob | |
dc.date.accessioned | 2021-12-23T16:10:37Z | - |
dc.date.available | 2021-12-23T16:10:37Z | - |
dc.date.issued | 2011 | |
dc.identifier.issn | 00032700 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/9320 | - |
dc.description.abstract | We have established a robust and versatile analytical platform for probing membrane protein function in a defined lipid environment on solid supports. This approach is based on vesicle capturing onto an ultrathin poly(ethylene glycol) (PEG) polymer brush functionalized with fatty acid moieties and subsequent vesicle fusion into a contiguous membrane. In order to ensure efficient formation of these tethered polymer-supported membranes (PSM), very small unilamellar vesicles (VSUV) containing fluorescent lipids or model transmembrane proteins were generated by detergent depletion with cyclodextrin. Thus, very rapid reconstitution of membrane proteins into PSM was possible in a format compatible with microfluidics. Moreover, surfaces could be regenerated with detergent solution and reused multiple times. Lipid and protein diffusion in these membranes was investigated by fluorescence recovery after photobleaching, single molecule tracking, and fluorescence correlation spectroscopy. Full mobility of lipids and a high degree of protein mobility as well as homogeneous diffusion of both were observed. Quantitative ligand binding studies by solid phase detection techniques confirmed functional integrity of a transmembrane receptor reconstituted into these PSM. Colocomotion of individual ligand-receptor complexes was detected, demonstrating the applicability for single molecule fluorescence techniques. | |
dc.description.sponsorship | DFGGerman Research Foundation (DFG)European Commission [PI405-4, PI405-5, SFB 944, PI405-3]; European CommunityEuropean Commission [223608]; FP6Marie-Curie EST fellowship [MEST-CT-2004-504272]; Minerva Foundation; We thank Gabriele Hikade and Hella Kenneweg for technical support, Jorg Nordmann and Markus Haase for the help with the dynamic light scattering, and Gotz Hofhaus for transmission electron microscopy. This project was supported by funding from the DFG (Grants PI405-4, PI405-5, and SFB 944) and by the European Community's Seventh Framework Programme (FP7/2007-2013) under Grant Agreement No. 223608 (IFN-action). J.P was supported by a Heisenberg Professorship from the DFG (Grant PI405-3). SW. was supported by a FP6Marie-Curie EST fellowship (Grant No. MEST-CT-2004-504272) and by a Ph.D. fellowship from the Minerva Foundation. | |
dc.language.iso | en | |
dc.publisher | AMER CHEMICAL SOC | |
dc.relation.ispartof | ANALYTICAL CHEMISTRY | |
dc.subject | 2D CRYSTALLIZATION | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Analytical | |
dc.subject | CONFORMATIONS | |
dc.subject | DYNAMICS | |
dc.subject | FLUORESCENCE CORRELATION SPECTROSCOPY | |
dc.subject | IMMOBILIZATION | |
dc.subject | LIPID-BILAYERS | |
dc.subject | MODEL MEMBRANES | |
dc.subject | PLANAR BILAYERS | |
dc.subject | SURFACE | |
dc.subject | TOTAL INTERNAL-REFLECTION | |
dc.title | Reconstitution of Membrane Proteins into Polymer-Supported Membranes for Probing Diffusion and Interactions by Single Molecule Techniques | |
dc.type | journal article | |
dc.identifier.doi | 10.1021/ac201294v | |
dc.identifier.isi | ISI:000294322100051 | |
dc.description.volume | 83 | |
dc.description.issue | 17 | |
dc.description.startpage | 6792 | |
dc.description.endpage | 6799 | |
dc.contributor.orcid | 0000-0002-6213-2872 | |
dc.contributor.researcherid | N-6093-2014 | |
dc.identifier.eissn | 15206882 | |
dc.publisher.place | 1155 16TH ST, NW, WASHINGTON, DC 20036 USA | |
dcterms.isPartOf.abbreviation | Anal. Chem. | |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | PiJa938 | - |