Journal Article

FZJ-2021-02672

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Structural Analysis of a Genetically Encoded FRET Biosensor by SAXS and MD Simulations

Reinartz, I. ; Sarter, M. ; Otten, J.FZJ* ; Höfig, H.FZJ* ; Pohl, M.FZJ* ; Schug, A.FZJ* ; Stadler, A. M.FZJ* ; Fitter, J. (Corresponding author)FZJ*

2021
MDPI Basel

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Please use a persistent id in citations: http://hdl.handle.net/2128/27997  doi:10.3390/s21124144

Abstract: 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.

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Contributing Institute(s):

1. Biotechnologie (IBG-1)
2. Zelluläre Strukturbiologie (IBI-6)
3. Neutronenstreuung (JCNS-1)
4. Neutronenstreuung und biologische Materie (IBI-8)

Research Program(s):

1. 2171 - Biological and environmental resources for sustainable use (POF4-217) (POF4-217)

Appears in the scientific report 2021

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