Black-lipid-membrane platform with diffusion suppression for confocal single-molecule FLIM-FRET analysis of gated membrane receptors and transporters (A01)

• Subject Area: Structural Biology ; Anatomy and Physiology ; Biophysics
• Term: from 2015 to 2019
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 258780946

G-protein coupled membrane receptors neurotensin-1 (NTSR-1) are expressed in bacteria, purified and labeled with two fluorophores for single-molecule Förster resonance energy transfer (smFRET). Reconstituted in specific liposomes, NTSR-1 or HCN2 ion channels will be fused with a planar black-lipid-membrane (BLM). Dynamic conformational changes are studied using time-resolved confocal as well as widefield smFRET microscopy in microfluidic chips with adjustable membrane potentials. Diffusion of membrane proteins in the BLM will be suppressed by an Annexin-V network for extended observation times.

• DFG Programme: CRC/Transregios
• Subproject of: TRR 166: High-end light microscopy elucidates membrane receptor function - ReceptorLight
• Major Instrumentation: Bilayer Explorer System
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Friedrich-Schiller-Universität Jena
• Project Head: Professor Dr. Michael Börsch