TY  - JOUR
A1  - Wawrzinek, Robert
A1  - Ziomkowska, Joanna
A1  - Heuveling, Johanna
A1  - Mertens, Monique
A1  - Herrmann, Andreas
A1  - Schneider, Erwin
A1  - Wessig, Pablo
T1  - DBD Dyes as Fluorescence Lifetime Probes to Study Conformational Changes
 in Proteins
JF  - CHEMISTRY-A EUROPEAN JOURNAL
N2  - Previously, [1,3]dioxolo[4,5-f][1,3]benzodioxole (DBD)-based fluorophores used as highly sensitive fluorescence lifetime probes reporting on their microenvironmental polarity have been described. Now, a new generation of DBD dyes has been developed. Although they are still sensitive to polarity, in contrast to the former DBD dyes, they have extraordinary spectroscopic properties even in aqueous surroundings. They are characterized by long fluorescence lifetimes (10-20ns), large Stokes shifts (approximate to 100nm), high photostabilities, and high quantum yields (>0.56). Here, the spectroscopic properties and synthesis of functionalized derivatives for labeling biological targets are described. Furthermore, thio-reactive maleimido derivatives of both DBD generations show strong intramolecular fluorescence quenching. This mechanism has been investigated and is found to undergo a photoelectron transfer (PET) process. After reaction with a thiol group, this fluorescence quenching is prevented, indicating successful bonding. Being sensitive to their environmental polarity, these compounds have been used as powerful fluorescence lifetime probes for the investigation of conformational changes in the maltose ATP-binding cassette transporter through fluorescence lifetime spectroscopy. The differing tendencies of the fluorescence lifetime change for both DBD dye generations promote their combination as a powerful toolkit for studying microenvironments in proteins.
KW  - dyes
KW  - pigments
KW  - electron transfer
KW  - fluorescent probes
KW  - maleimides
KW  - MalF
KW  - photoelectron transfer
Y1  - 2013
U6  - https://doi.org/10.1002/chem.201302368
SN  - 0947-6539
SN  - 1521-3765
VL  - 19
IS  - 51
SP  - 17349
EP  - 17357
PB  - WILEY-V C H VERLAG GMBH
CY  - WEINHEIM
ER  -