TY  - JOUR
A1  - Kienzler, Andrea Altevogt Nee
A1  - Flehr, Roman
A1  - Gehne, Sören
A1  - Kumke, Michael Uwe
A1  - Bannwarth, Willi
T1  - Verification and biophysical characterization of a New Three-Color Forster Resonance-Energy-Transfer (FRET) System in DNA
JF  - Helvetica chimica acta
N2  - We report on a new three-color FRET system consisting of three fluorescent dyes, i.e., of a carbostyril (=quinolin-2(1H)-one)-derived donor D, a (bathophenanthroline)ruthenium complex as a relay chromophore A1, and a Cy dye as A2 (FRET=Forster resonance-energy-transfer) (cf. Fig. 1). With their widely matching spectroscopic properties (cf. Fig. 2), the combination of these dyes yielded excellent FRET efficiencies. Furthermore, fluorescence lifetime measurements revealed that the long fluorescence lifetime of the Ru complex was transferred to the Cy dye offering the possibility to measure the whole system in a time-resolved mode. The FRET system was established on double-stranded DNA (cf. Fig. 3) but it should also be generally applicable to other biomolecules.
KW  - Forster resonance energy transfer (FRET) system
KW  - DNA
KW  - Fluorescence
KW  - Ruthenium complexes
Y1  - 2012
U6  - https://doi.org/10.1002/hlca.201100460
SN  - 0018-019X
VL  - 95
IS  - 4
SP  - 543
EP  - 555
PB  - Wiley-VCH
CY  - Weinheim
ER  - 
TY  - JOUR
A1  - Gehne, Sören
A1  - Flehr, Roman
A1  - Kienzler, Andrea Altevogt Nee
A1  - Berg, Maik
A1  - Bannwarth, Willi
A1  - Kumke, Michael Uwe
T1  - Dye dynamics in three-color FRET samples
JF  - The journal of physical chemistry : B, Condensed matter, materials, surfaces, interfaces & biophysical chemistry
N2  - Time-resolved emission data (fluorescence decay and fluorescence depolarization) of two three-color Forster resonance energy transfer (tc-FRET) systems consisting of a carbostyril donor (D), a ruthenium complex (Ru) as relay dye, and a Cy5 derivative (Cy) or, optionally, an anthraquinone quencher (Q) were carefully analyzed using advanced distribution analysis models. Thereby, it is possible to get information on the flexibility and mobility of the chromophores which are bound to double stranded (ds) DNA. Especially the distance distribution based on the analysis of the fluorescence depolarization is an attractive approach to complement data of fluorescence decay time analysis. The distance distributions extracted from the experimental data were in excellent agreement with those determined from accessible volume (AV) simulations. Moreover, the study showed that for tc-FRET systems the combination of dyes emitting on different time scales (e.g., nanoseconds vs microseconds) is highly beneficial in the distribution analysis of time-resolved luminescence data in cases where macromolecules such as DNA are involved. Here, the short lifetimes can yield information on the rotation of the dye molecule itself and the long lifetime can give insight in the overall dynamics of the macromolecule.
Y1  - 2012
U6  - https://doi.org/10.1021/jp3064273
SN  - 1520-6106
VL  - 116
IS  - 35
SP  - 10798
EP  - 10806
PB  - American Chemical Society
CY  - Washington
ER  -