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@ARTICLE{Reinartz:893282,
author = {Reinartz, Ines and Sarter, Mona and Otten, Julia and
Höfig, Henning and Pohl, Martina and Schug, Alexander and
Stadler, Andreas M. and Fitter, Jörg},
title = {{S}tructural {A}nalysis of a {G}enetically {E}ncoded {FRET}
{B}iosensor by {SAXS} and {MD} {S}imulations},
journal = {Sensors},
volume = {21},
number = {12},
issn = {1424-8220},
address = {Basel},
publisher = {MDPI},
reportid = {FZJ-2021-02672},
pages = {4144 -},
year = {2021},
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.},
cin = {IBG-1 / IBI-6 / JCNS-1 / IBI-8},
ddc = {620},
cid = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)IBI-6-20200312 /
I:(DE-Juel1)JCNS-1-20110106 / I:(DE-Juel1)IBI-8-20200312},
pnm = {2171 - Biological and environmental resources for
sustainable use (POF4-217)},
pid = {G:(DE-HGF)POF4-2171},
typ = {PUB:(DE-HGF)16},
pubmed = {34208740},
UT = {WOS:000667410700001},
doi = {10.3390/s21124144},
url = {https://juser.fz-juelich.de/record/893282},
}