Genetically Encoded Förster Resonance Energy Transfer-Based Biosensors Studied on the Single-Molecule Level

Höfig, Henning; Fitter, Joerg; Otten, Julia; Pohl, Martina; Steffen, Victoria; Boersma, Arnold J.

doi:10.1021/acssensors.8b00143

Journal Article

FZJ-2018-04316

Genetically Encoded Förster Resonance Energy Transfer-Based Biosensors Studied on the Single-Molecule Level

Höfig, H.FZJ* ; Otten, J.FZJ* ; Steffen, V.FZJ* ; Pohl, M.FZJ* ; Boersma, A. J. ; Fitter, J. (Corresponding author)FZJ*

2018
ACS Publications Washington, DC

ACS sensors 3(8), 1462–1470 (2018) [10.1021/acssensors.8b00143]

This record in other databases:

Please use a persistent id in citations: doi:10.1021/acssensors.8b00143

Abstract: 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.

Classification: