title: Rapid transfer of transmembrane proteins for single molecule dimerization assays in polymer-supported membranes.
abstract: Dimerization of transmembrane receptors is a key regulatory factor in cellular communication, which has remained challenging to study under well-defined conditions in vitro. We developed a novel strategy to explore membrane protein interactions in a controlled lipid environment requiring minute sample quantities. By rapid transfer of transmembrane proteins from mammalian cells into polymer-supported membranes, membrane proteins could be efficiently fluorescence labeled and reconstituted with very low background. Thus, differential ligand-induced dimerization of the type I interferon (IFN) receptor subunits IFNAR1 and IFNAR2 could be probed quantitatively at physiologically relevant concentrations by single molecule imaging. These measurements clearly support a regulatory role of the affinity of IFNs toward IFNAR1 for controlling the level of receptor dimerization.
authors.affiliation: Division of Biophysics, Department of Biology, University of Osnabrück , Barbarastr. 11, 49076 Osnabrück, Germany.
authors.forename: Friedrich
authors.initials: F
authors.lastname: Roder
authors.forename: Stephan
authors.initials: S
authors.lastname: Wilmes
authors.forename: Christian P
authors.initials: CP
authors.lastname: Richter
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
mesh_terms: D000818:Animals; D019281:Dimerization; D008024:Ligands; D008322:Mammals; D008565:Membrane Proteins / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*
chemical_list: D008024:Ligands; D008565:Membrane Proteins; D011108:Polymers