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@ARTICLE{Hfig:857784,
author = {Höfig, Henning and Cerminara, Michele and Ritter, Ilona
and Schöne, Antonie and Pohl, Martina and Steffen, Victoria
and Walter, Julia and Vergara Dal Pont, Ignacio and
Katranidis, Alexandros and Fitter, Jörg},
title = {{S}ingle-{M}olecule {S}tudies on a {FRET} {B}iosensor:
{L}essons from a {C}omparison of {F}luorescent {P}rotein
{E}quipped versus {D}ye-{L}abeled {S}pecies},
journal = {Molecules},
volume = {23},
number = {12},
issn = {1420-3049},
address = {Basel},
publisher = {MDPI},
reportid = {FZJ-2018-06751},
pages = {3105 -},
year = {2018},
abstract = {Bacterial periplasmic binding proteins (PBPs) undergo a
pronounced ligand-induced conformational change which can be
employed to monitor ligand concentrations. The most common
strategy to take advantage of this conformational change for
a biosensor design is to use a Förster resonance energy
transfer (FRET) signal. This can be achieved by attaching
either two fluorescent proteins (FPs) or two organic
fluorescent dyes of different colors to the PBPs in order to
obtain an optical readout signal which is closely related to
the ligand concentration. In this study we compare a
FP-equipped and a dye-labeled version of the
glucose/galactose binding protein MglB at the
single-molecule level. The comparison demonstrates that
changes in the FRET signal upon glucose binding are more
pronounced for the FP-equipped sensor construct as compared
to the dye-labeled analog. Moreover, the FP-equipped sensor
showed a strong increase of the FRET signal under crowding
conditions whereas the dye-labeled sensor was not influenced
by crowding. The choice of a labeling scheme should
therefore be made depending on the application of a
FRET-based sensor.},
cin = {ICS-5 / IBT-1},
ddc = {540},
cid = {I:(DE-Juel1)ICS-5-20110106 / I:(DE-Juel1)VDB55},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551)},
pid = {G:(DE-HGF)POF3-551},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30486450},
UT = {WOS:000454523000055},
doi = {10.3390/molecules23123105},
url = {https://juser.fz-juelich.de/record/857784},
}