@article{WessigWawrzinekMoellnitzetal.2011,
  author    = {Wessig, Pablo and Wawrzinek, Robert and Moellnitz, Kristian and Feldbusch, Elvira and Schilde, Uwe},
  title     = {A new class of fluorescent dyes based on 1,3-benzodioxole and [1,3]-dioxolo[4.5-f]benzodioxole},
  series = {Tetrahedron letters},
  volume    = {52},
  journal   = {Tetrahedron letters},
  number    = {46},
  publisher = {Elsevier},
  address   = {Oxford},
  issn      = {0040-4039},
  doi       = {10.1016/j.tetlet.2011.09.058},
  pages     = {6192 -- 6195},
  year      = {2011},
  abstract  = {We report on synthesis and photophysical properties of a new class of fluorescent dyes. They are characterized by large Stokes-shifts, long fluorescence lifetimes in organic solvents and a pronounced dependency of the fluorescence lifetime on the solvent polarity. Also worthy of note is the high bleaching stability. To provide access to biochemical and medical applications a series of derivatives were prepared, which exhibit specific reactivity towards different biologically relevant functional groups (carboxylic acids, amines, maleimides, N-hydroxysuccinimide esters). Furthermore, two alkynes were prepared, which could be used in 'Click' chemistry.},
  language  = {en}
}
@article{WawrzinekZiomkowskaHeuvelingetal.2013,
  author    = {Wawrzinek, Robert and Ziomkowska, Joanna and Heuveling, Johanna and Mertens, Monique and Herrmann, Andreas and Schneider, Erwin and Wessig, Pablo},
  title     = {DBD Dyes as Fluorescence Lifetime Probes to Study Conformational Changes in Proteins},
  series = {CHEMISTRY-A EUROPEAN JOURNAL},
  volume    = {19},
  journal   = {CHEMISTRY-A EUROPEAN JOURNAL},
  number    = {51},
  publisher = {WILEY-V C H VERLAG GMBH},
  address   = {WEINHEIM},
  issn      = {0947-6539},
  doi       = {10.1002/chem.201302368},
  pages     = {17349 -- 17357},
  year      = {2013},
  abstract  = {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.},
  language  = {en}
}
@article{WawrzinekWessigMoellnitzetal.2012,
  author    = {Wawrzinek, Robert and Wessig, Pablo and M{\"o}llnitz, Kristian and Nikolaus, Joerg and Schwarzer, Roland and M{\"u}ller, Peter and Herrmann, Andreas},
  title     = {DBD dyes as fluorescent probes for sensing lipophilic environments},
  series = {Bioorganic \& medicinal chemistry letters : a Tetrahedron publication for rapid dissemination of preliminary communications on all aspects of bioorganic chemistry, medicinal chemistry and related disciplines},
  volume    = {22},
  journal   = {Bioorganic \& medicinal chemistry letters : a Tetrahedron publication for rapid dissemination of preliminary communications on all aspects of bioorganic chemistry, medicinal chemistry and related disciplines},
  number    = {17},
  publisher = {Elsevier},
  address   = {Oxford},
  issn      = {0960-894X},
  doi       = {10.1016/j.bmcl.2012.07.056},
  pages     = {5367 -- 5371},
  year      = {2012},
  abstract  = {Small fluorescent organic molecules based on [1,3]dioxolo[4,5-f][1,3]benzodioxole (DBD) could be used as probes for lipophillic microenvironments in aqueous solutions by indicating the critical micelles concentration of detergents and staining cell organelles. Their fluorescence lifetime decreases drastically by the amount of water in their direct environment. Therefore they are potential probes for fluorescence lifetime imaging microscopy (FLIM).},
  language  = {en}
}
@article{MeynersWawrzinekKraemeretal.2014,
  author    = {Meyners, Christian and Wawrzinek, Robert and Kraemer, Andreas and Hinz, Steffen and Wessig, Pablo and Meyer-Almes, Franz-Josef},
  title     = {A fluorescence lifetime-based binding assay for acetylpolyamine amidohydrolases from Pseudomonas aeruginosa using a [1,3]dioxolo[4,5-f][1,3]benzodioxole (DBD) ligand probe},
  series = {Analytical \& bioanalytical chemistry},
  volume    = {406},
  journal   = {Analytical \& bioanalytical chemistry},
  number    = {20},
  publisher = {Springer},
  address   = {Heidelberg},
  issn      = {1618-2642},
  doi       = {10.1007/s00216-014-7886-5},
  pages     = {4889 -- 4897},
  year      = {2014},
  abstract  = {High-throughput assays for drug screening applications have to fulfill particular specifications. Besides the capability to identify even compounds with low potency, one of the major issues is to minimize the number of false-positive hits in a screening campaign in order to reduce the logistic effort for the subsequent cherry picking and confirmation procedure. In this respect, fluorescence lifetime (FLT) appears as an ideal readout parameter that is supposed to be robust against autofluorescent and light-absorbing compounds, the most common source of systematic false positives. The extraordinary fluorescence features of the recently discovered [1,3]dioxolo[4,5-f][1,3]benzodioxole dyes were exploited to develop an FLT-based binding assay with exceptionally robust readout. The assay setup was comprehensively validated and shown to comply not only with all requirements for a powerful high-throughput screening assay but also to be suitable to determine accurate binding constants for inhibitors against enzymes of the histone deacetylase family. Using the described binding assay, the first inhibitors against three members of this enzyme family from Pseudomonas aeruginosa were identified. The compounds were characterized in terms of potency and selectivity profile. The novel ligand probe should also be applicable to other homologues of the histone deacetylase family that are inhibited by N-hydroxy-N'-phenyloctandiamide.},
  language  = {en}
}
@article{HeuvelingFrochauxZiomkowskaetal.2014,
  author    = {Heuveling, Johanna and Frochaux, Violette and Ziomkowska, Joanna and Wawrzinek, Robert and Wessig, Pablo and Herrmann, Andreas and Schneider, Erwin},
  title     = {Conformational changes of the bacterial type I ATP-binding cassette importer HisQMP(2) at distinct steps of the catalytic cycle},
  series = {Biochimica et biophysica acta : Biomembranes},
  volume    = {1838},
  journal   = {Biochimica et biophysica acta : Biomembranes},
  number    = {1},
  publisher = {Elsevier},
  address   = {Amsterdam},
  issn      = {0005-2736},
  doi       = {10.1016/j.bbamem.2013.08.024},
  pages     = {106 -- 116},
  year      = {2014},
  abstract  = {Prokaryotic solute binding protein-dependent ATP-binding cassette import systems are divided into type land type II and mechanistic differences in the transport process going along with this classification are under intensive investigation. Little is known about the conformational dynamics during the catalytic cycle especially concerning the transmembrane domains. The type I transporter for positively charged amino acids from Salmonella enterica serovar Typhimurium (1A0-Hi5QMP2) was studied by limited proteolysis in detergent solution in the absence and presence of co-factors including ATP, ADP, LAO/arginine, and Mg2+ ions. Stable peptide fragments could be obtained and differentially susceptible cleavage sites were determined by mass spectrometry as Lys-258 in the nucleotide-binding subunit, HisP, and Arg-217/Arg-218 in the transmembrane subunit, HisQ In contrast, transmembrane subunit HisM was gradually degraded but no stable fragment could be detected. HisP and HisQ were equally resistant under pre- and post-hydrolysis conditions in the presence of arginine-loaded solute-binding protein LAO and ATP/ADP. Some protection was also observed with LAO/arginine alone, thus reflecting binding to the transporter in the apo-state and transmembrane signaling. Comparable digestion patterns were obtained with the transporter reconstituted into proteoliposomes and nanodiscs. Fluorescence lifetime spectroscopy confirmed the change of HisQ(R218) to a more apolar microenvironment upon ATP binding and hydrolysis. Limited proteolysis was subsequently used as a tool to study the consequences of mutations on the transport cycle. Together, our data suggest similar conformational changes during the transport cycle as described for the maltose ABC transporter of Escherichia coli, despite distinct structural differences between both systems.},
  language  = {en}
}