TY  - JOUR
AU  - Reinartz, Ines
AU  - Sarter, Mona
AU  - Otten, Julia
AU  - Höfig, Henning
AU  - Pohl, Martina
AU  - Schug, Alexander
AU  - Stadler, Andreas M.
AU  - Fitter, Jörg
TI  - Structural Analysis of a Genetically Encoded FRET Biosensor by SAXS and MD Simulations
JO  - Sensors
VL  - 21
IS  - 12
SN  - 1424-8220
CY  - Basel
PB  - MDPI
M1  - FZJ-2021-02672
SP  - 4144 -
PY  - 2021
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 34208740
UR  - <Go to ISI:>//WOS:000667410700001
DO  - DOI:10.3390/s21124144
UR  - https://juser.fz-juelich.de/record/893282
ER  -