TY  - JOUR
AU  - Höfig, Henning
AU  - Otten, Julia
AU  - Steffen, Victoria
AU  - Pohl, Martina
AU  - Boersma, Arnold J.
AU  - Fitter, Joerg
TI  - Genetically Encoded Förster Resonance Energy Transfer-Based Biosensors Studied on the Single-Molecule Level
JO  - ACS sensors
VL  - 3
IS  - 8
SN  - 2379-3694
CY  - Washington, DC
PB  - ACS Publications
M1  - FZJ-2018-04316
SP  - 1462–1470
PY  - 2018
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:29979038
UR  - <Go to ISI:>//WOS:000443103800006
DO  - DOI:10.1021/acssensors.8b00143
UR  - https://juser.fz-juelich.de/record/850274
ER  -