Publication details

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.

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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