BIOPHYSICAL STUDY OF THE INTERACTION OF ZINC IONS WITH DNA
| Autoři | |
|---|---|
| Rok publikování | 2013 |
| Druh | Článek ve sborníku |
| Konference | XIII. Pracovní setkání fyzikálních chemiků a elektrochemiků, sborník |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Fyzikální chemie a teoretická chemie |
| Klíčová slova | DNA;zinc;UV/VIS spectrophotometry |
| Popis | We focused interactions of zinc ions and DNA in this study. Interactions were monitored using UV/VIS spectrophotometry and gel electrophoresis. Firstly, we isolated and amplified fragment of DNA of 498 bp. DNA was subsequently incubated with Zn(II) ions for 60 min at 25 C. After incubation, samples were dialyzed and analysed immediately. In this way, DNA was converted into a metal bound DNA (M-DNA). Interaction of Zn(II) with DNA showed a change of the absorption spectrum (190-350 nm) and a decrease in the M-DNA melting temperature. Spectrophotometric (UV/VIS) analysis showed that increasing concentrations of Zn(II) led to an increase in absorbance at 200 nm and a decrease in absorbance at 260 nm. Application of 5.5 microM Zn(II) caused a decrease in the melting temperature (Tm) for about 7.5 C in average in comparison with control (75.5 +- 3 C). The lowest melting temperature (60.5 +- 2.5 C) was observed after application of 33 microM Zn(II). On the other hand, application of two concentrations, 44 and 55 microM Zn(II), caused no changes in Tm. Gel electrophoresis proved significance of Zn(II) ions in the DNA renaturation. Samples of M-DNA (5microg/ml DNA + 5.5-55microM Zn(II) showed a significant changes in the renaturation in comparison with control (5microg/ml DNA). This study has proved that the zinc ion can incorporate into DNA (M-DNA) at standard conditions (25 C). This work suggests that the new structure of DNA (M-DNA) may occur in living organisms. |