Rapid Transfer of Transmembrane Proteins for Single Molecule Dimerization Assays in Polymer-Supported Membranes

Autor(en): Roder, Friedrich
Wilmes, Stephan
Richter, Christian P.
Piehler, Jacob 
Stichwörter: ACTIVATION; ARCHITECTURE; Biochemistry & Molecular Biology; DIFFUSION; I INTERFERON RECEPTOR; LIPIDS; ORGANIZATION; PLASMA-MEMBRANE; RECOGNITION; RECONSTITUTION; REVEALS
Erscheinungsdatum: 2014
Herausgeber: AMER CHEMICAL SOC
Journal: ACS CHEMICAL BIOLOGY
Volumen: 9
Ausgabe: 11
Startseite: 2479
Seitenende: 2484
Zusammenfassung: 
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.
ISSN: 15548929
DOI: 10.1021/cb5005806

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