1 research outputs found
Dye Dynamics in Three-Color FRET Samples
Time-resolved emission data (fluorescence decay and fluorescence
depolarization) of two three-color FoĢrster 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