Description: Single-Molecule Studies on a FRET Biosensor: Lessons from a Comparison of Fluorescent Protein Equipped versus Dye-Labeled Species

This title appears in the Scientific Report : 2018

Single-Molecule Studies on a FRET Biosensor: Lessons from a Comparison of Fluorescent Protein Equipped versus Dye-Labeled Species

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 transfe...

Personal Name(s):	Höfig, Henning
	Cerminara, Michele / Ritter, Ilona / Schöne, Antonie / Pohl, Martina / Steffen, Victoria / Walter, Julia / Vergara Dal Pont, Ignacio / Katranidis, Alexandros / Fitter, Jörg (Corresponding author)
Contributing Institute:	Molekulare Biophysik; ICS-5 Biotechnologie 1; IBT-1
Published in:	Molecules, 23 (2018) 12, S. 3105 -
Imprint:	Basel MDPI 2018
PubMed ID:	30486450
DOI:	10.3390/molecules23123105
Document Type:	Journal Article
Research Program:	Functional Macromolecules and Complexes
Link:	Get full text Get full text OpenAccess OpenAccess
	Publikationsportal JuSER

Please use the identifier: http://dx.doi.org/10.3390/molecules23123105 in citations.
Please use the identifier: http://hdl.handle.net/2128/20695 in citations.

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.