%0 Journal Article
%A Höfig, Henning
%A Otten, Julia
%A Steffen, Victoria
%A Pohl, Martina
%A Boersma, Arnold J.
%A Fitter, Joerg
%T Genetically Encoded Förster Resonance Energy Transfer-Based Biosensors Studied on the Single-Molecule Level
%J ACS sensors
%V 3
%N 8
%@ 2379-3694
%C Washington, DC
%I ACS Publications
%M FZJ-2018-04316
%P 1462–1470
%D 2018
%X Genetically encoded Förster resonance energy transfer (FRET)-based biosensors for the quantification of ligand molecules change the magnitude of FRET between two fluorescent proteins upon binding a target metabolite. When highly sensitive sensors are being designed, extensive sensor optimization is essential. However, it is often difficult to verify the ideas of modifications made to a sensor during the sensor optimization process because of the limited information content of ensemble FRET measurements. In contrast, single-molecule detection provides detailed information and higher accuracy. Here, we investigated a set of glucose and crowding sensors on the single-molecule level. We report the first comprehensive single-molecule study of FRET-based biosensors with reasonable counting statistics and identify characteristics in the single-molecule FRET histograms that constitute fingerprints of sensor performance. Hence, our single-molecule approach extends the toolbox of methods aiming to understand and optimize the design of FRET-based biosensors.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:29979038
%U <Go to ISI:>//WOS:000443103800006
%R 10.1021/acssensors.8b00143
%U https://juser.fz-juelich.de/record/850274