%0 Journal Article
%A Reinartz, Ines
%A Sarter, Mona
%A Otten, Julia
%A Höfig, Henning
%A Pohl, Martina
%A Schug, Alexander
%A Stadler, Andreas M.
%A Fitter, Jörg
%T Structural Analysis of a Genetically Encoded FRET Biosensor by SAXS and MD Simulations
%J Sensors
%V 21
%N 12
%@ 1424-8220
%C Basel
%I MDPI
%M FZJ-2021-02672
%P 4144 -
%D 2021
%X Inspired by the modular architecture of natural signaling proteins, ligand binding proteins are equipped with two fluorescent proteins (FPs) in order to obtain Förster resonance energy transfer (FRET)-based biosensors. Here, we investigated a glucose sensor where the donor and acceptor FPs were attached to a glucose binding protein using a variety of different linker sequences. For three resulting sensor constructs the corresponding glucose induced conformational changes were measured by small angle X-ray scattering (SAXS) and compared to recently published single molecule FRET results (Höfig et al., ACS Sensors, 2018). For one construct which exhibits a high change in energy transfer and a large change of the radius of gyration upon ligand binding, we performed coarse-grained molecular dynamics simulations for the ligand-free and the ligand-bound state. Our analysis indicates that a carefully designed attachment of the donor FP is crucial for the proper transfer of the glucose induced conformational change of the glucose binding protein into a well pronounced FRET signal change as measured in this sensor construct. Since the other FP (acceptor) does not experience such a glucose induced alteration, it becomes apparent that only one of the FPs needs to have a well-adjusted attachment to the glucose binding protein.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ 34208740
%U <Go to ISI:>//WOS:000667410700001
%R 10.3390/s21124144
%U https://juser.fz-juelich.de/record/893282