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BP: Fachverband Biologische Physik

BP 19: Poster VII

BP 19.25: Poster

Dienstag, 17. März 2020, 14:00–16:00, P2/3OG

A proof-of-principle set-up linking ion channel conformation and function by combining single-molecule FRET and single-channel recording — •Steven Vanuytsel, Christopher Parperis, and Mark Wallace — Department of Chemistry, King's College London, London, UK

Building a bridge between static high-resolution membrane protein structures and the kinetic information obtained via patch-clamping remains challenging due to significant incompatibilities in the experimental conditions required to measure these two signals. Nevertheless, when successful, such a method would provide a plethora of information by allowing the construction of a molecular movie that links function to conformation.

Optical single-channel recording (oSCR) shifts the measurement of ionic current from an electrical to an optical readout, allowing parallel interrogation of multiple channels simultaneously. Furthermore, shifting from an electrical to an optical set-up allows a straightforward route to measure function and conformation in parallel by interrogating the latter using single-molecule Förster Resonance Energy Transfer (smFRET).

Here, we report on our recent work to establish parallelized smFRET and oSCR recordings in a proof-of-principle set-up that allows control over single-molecule *gating* kinetics of a protein nanopore that stochastically senses fluorescently labelled DNA, thereby allowing us to record well-controlled simultaneous events.