Electrochemical and capillary zone electrophoresis study of pyrimidine residues modification by osmium tetroxide, 2,2´-bipyridine. Detection of unpaired and mispaired thymines in DNA heteroduplexes.

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Authors

FOJTA Miroslav KOSTEČKA Pavel BITTOVÁ Miroslava BARTOŠÍK Martin HIANIK Tibor TRNKOVÁ Libuše PALEČEK Emil

Year of publication 2009
Type Article in Proceedings
Conference XXIX. moderní elektrochemické metody - sborník příspěvků
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords DNA; OsO4 bipy; voltammetry; capillary electrophoresis
Description In this work we used capillary zone electrophoresis (CZE) in combination with square wave voltammetry (SWV) at a pyrolytic graphite electrode (PGE) to study reactivity of different pyrimidine residues, thymine (T), uracil (U), cytosine (C) and 5 methyl cytosine (meC), within a pentanucleotide AAAAY (where Y stands for the pyrimidine residue), towards Os,bipy. Since adenine practically does not react with Os,L, Y residues represent the only reactive centre within the pentamers and their reactivity can thus be easily compared. Both techniques revealed the order of the Y reactivities as follows: T>>MeC~U>C. Hence, our results confirmed (a) highly preferential modification of T among all Ys and (b) increased reactivity of MeC, compared to C that was reported by other authors. However, despite recent papers presenting Os,L modification as useful tool for MeC detection in DNA, the strong modification of T within natural DNA (where T is much more abundant than MeC) makes this approach hardly applicable for real analyses of DNA methylation.
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