Berlin 2018 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 23: Bioimaging and Biopspectroscopy II
BP 23.3: Vortrag
Mittwoch, 14. März 2018, 15:30–15:45, H 2013
Enhanced fluorescence resonance energy transfer in G protein-coupled receptor probes by nano-coated microscopy coverslips — •Benjamin Schreiber1, Michael Kauk1,2,4, Hannah S Heil1, Martin Kamp3, Sven Höfling3, Carsten Hoffmann1,2,4, and Katrin G Heinze1 — 1Rudolf Virchow Zentrum, Universität Würzburg — 2Institut für Pharmakologie und Toxikologie, Universität Würzburg — 3Technische Physik, Universität Würzburg — 4Institut für Molekulare Zellbiologie, Universität Jena
For probing biomolecular interactions in a live-cell setting the distance depending Fluorescence resonance energy transfer (FRET) is often the method of choice. G-protein-coupled receptors mediates cellular responses and communication across cellular membranes, and is the largest known class of molecular targets with proven therapeutic value. The design of FRET probes is crucial to ensure unhampered functionality and binding kinetics of the molecular complex. Thus, such FRET probes usually require labeling compromises with limited FRET efficiencies. Here, we present an approach to optimize the energy transfer without changing the design of the FRET probe. We show that gold coated glass cover slips allow reinforcing the otherwise forbidden donor-acceptor energy transfer by virtual optimization of the dipole orientation. We show resulting enhanced FRET on our nano-coatings for the ligand driven activation of M1 muscarinic acetylcholine receptors labeled with a CFP-FlAsH pair. We believe that our techniques has particular potential for pharmaceutical drug screening.