Affiliation: | 1. The Czech Academy of Sciences, Institute of Biophysics, Královopolská 135, 612 65 Brno, Czech Republic
These authors contributed equally to this work.;2. Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo náměstí 2, 16610 Prague 6, Czech Republic
Department of Organic Chemistry, Faculty of Science, Charles University in Prague, Hlavova 8, Prague-2, 12843 Czech Republic
These authors contributed equally to this work.;3. Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo náměstí 2, 16610 Prague 6, Czech Republic;4. The Czech Academy of Sciences, Institute of Biophysics, Královopolská 135, 612 65 Brno, Czech Republic;5. The Czech Academy of Sciences, Institute of Biophysics, Královopolská 135, 612 65 Brno, Czech Republic
Central European Institute of Technology, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic |
Abstract: | Six-valent osmium (osmate) complexes with nitrogenous ligands have previously been used for the modification and redox labeling of biomolecules involving vicinal diol moieties (typically, saccharides or RNA). In this work, aliphatic (3,4-dihydroxybutyl and 3,4-dihydroxybut-1-ynyl) or cyclic (6-oxo-6-(cis-3,4-dihydroxypyrrolidin-1-yl)hex-2-yn-1-yl, PDI) vicinal diols are attached to nucleobases to functionalize DNA for subsequent redox labeling with osmium(VI) complexes. The diol-linked 2′-deoxyribonucleoside triphosphates were used for the polymerase synthesis of diol-linked DNA, which, upon treatment with K2OsO3 and bidentate nitrogen ligands, gave the desired Os-labeled DNA, which were characterized by means of the gel-shift assay and ESI-MS. Through ex situ square-wave voltammetry at a basal plane pyrolytic graphite electrode, the efficiency of modification/labeling of individual diols was evaluated. The results show that the cyclic cis-diol (PDI) was a better target for osmylation than that of the flexible aliphatic ones (alkyl- or alkynyl-linked). The osmate adduct-specific voltammetric signal obtained for OsVI-treated DNA decorated with PDI showed good proportionality to the number of PDI per DNA molecule. The OsVI reagents (unlike OsO4) do not attack nucleobases; thus offering specificity of modification on the introduced glycol targets. |