Electrochemistry of epigenetic modulator zebularine:
Voltammetric determination in cell culturing media and
monitoring of polymerase incorporation into DNA

J 2026

Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA

DAŇHEL, Aleš; Peter ŠEBEST; Daniel DOBROVODSKÝ; Marek ZELINKA; Pavlina HAVRANOVA et al.

Základní údaje

Originální název

Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA

Autoři

DAŇHEL, Aleš; Peter ŠEBEST; Daniel DOBROVODSKÝ; Marek ZELINKA ; Pavlina HAVRANOVA; Luděk HAVRAN; Miloslava FOJTOVÁ a Miroslav FOJTA

Vydání

ELECTROCHIMICA ACTA, OXFORD, PERGAMON-ELSEVIER SCIENCE LTD, 2026, 0013-4686

Další údaje

Typ výsledku

Článek v odborném periodiku

Stát vydavatele

Velká Británie a Severní Irsko

Impakt faktor

Impact factor: 5.600 v roce 2024

Označené pro přenos do RIV

DOI

https://doi.org/10.1016/j.electacta.2026.148707

UT WoS

001730948700001

Klíčová slova anglicky

DNA; Mercury electrode; Silver solid amalgam; Pyrolytic graphite; Voltammetry; Zebularine; 5-aza cytidine; Polymerase incorporation

Změněno: 27. 5. 2026 10:40, Mgr. Marek Zelinka

Anotace

V originále

Electrochemical properties of epigenetic modulator zebularine (Zeb) were studied for the first time using cyclic voltammetry (CV) at three types of working electrodes (hanging mercury drop electrode - HMDE; mercury meniscus-modified silver solid amalgam electrode - m-AgSAE; and basal plane pyrolytic graphite electrode - bPGE). CV responses of Zeb were investigated within the pH range from 2 to 12. Depending on the electrode type, Zeb gave up to three cathodic peaks ("c1", "c2", "c3") and an anodic peak "a" corresponding to oxidation of the product of Zeb reduction. Using optimized pretreatment of the solid electrodes, concentration dependences of the peak c1 were measured in ammonium formate, phosphate buffer pH 6.9 at the m-AgSAE, or in sodium acetate buffer pH 5.0 at the bPGE. LOD of 5 & micro;M was achieved with all three electrodes in the buffer solutions. Using HMDE, the same LOD was attained also in cell culturing media without any sample pretreatment. Incorporation of 2 '-deoxyzebularine 5 '-triphosphate into DNA was studied using primer extension and terminal deoxynucleotidyl transferase tailing techniques. Incorporation of Zeb exhibited poor efficacy; nevertheless, products of TdT tailing with deoxyZeb were successfully detected by ex situ CV. Interestingly, another epigenetic modulator, 5-azacytidine showed facile incorporation in both approaches. These observations were in accord with the presumably different mechanism of action of the two epigenetic drugs in cells. We demonstrate usefulness of a simple electrochemical method to assess different facileness of polymerase incorporation of two epigenetic drugs, cytidine analogues Zeb and 5-aC, by DNA polymerases.

Citovat

DAŇHEL, Aleš; Peter ŠEBEST; Daniel DOBROVODSKÝ; Marek ZELINKA; Pavlina HAVRANOVA; Luděk HAVRAN; Miloslava FOJTOVÁ a Miroslav FOJTA. Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA. ELECTROCHIMICA ACTA. OXFORD: PERGAMON-ELSEVIER SCIENCE LTD, 2026, roč. 563, s. 1-10. ISSN 0013-4686. Dostupné z: https://doi.org/10.1016/j.electacta.2026.148707.

@article{2575220,
   author = {Daňhel, Aleš and Šebest, Peter and Dobrovodský, Daniel and Zelinka, Marek and Havranova, Pavlina and Havran, Luděk and Fojtová, Miloslava and Fojta, Miroslav},
   article_location = {OXFORD},
   doi = {https://doi.org/10.1016/j.electacta.2026.148707},
   keywords = {DNA; Mercury electrode; Silver solid amalgam; Pyrolytic graphite; Voltammetry; Zebularine; 5-aza cytidine; Polymerase incorporation},
   issn = {0013-4686},
   journal = {ELECTROCHIMICA ACTA},
   title = {Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA},
   volume = {563},
   year = {2026}
}

TY  - JOUR
ID  - 2575220
AU  - Daňhel, Aleš - Šebest, Peter - Dobrovodský, Daniel - Zelinka, Marek - Havranova, Pavlina - Havran, Luděk - Fojtová, Miloslava - Fojta, Miroslav
PY  - 2026
TI  - Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA
JF  - ELECTROCHIMICA ACTA
VL  - 563
SP  - 1-10
EP  - 1-10
PB  - PERGAMON-ELSEVIER SCIENCE LTD
SN  - 00134686
KW  - DNA
KW  - Mercury electrode
KW  - Silver solid amalgam
KW  - Pyrolytic graphite
KW  - Voltammetry
KW  - Zebularine
KW  - 5-aza cytidine
KW  - Polymerase incorporation
N2  - Electrochemical properties of epigenetic modulator zebularine (Zeb) were studied for the first time using cyclic voltammetry (CV) at three types of working electrodes (hanging mercury drop electrode - HMDE; mercury meniscus-modified silver solid amalgam electrode - m-AgSAE; and basal plane pyrolytic graphite electrode - bPGE). CV responses of Zeb were investigated within the pH range from 2 to 12. Depending on the electrode type, Zeb gave up to three cathodic peaks ("c1", "c2", "c3") and an anodic peak "a" corresponding to oxidation of the product of Zeb reduction. Using optimized pretreatment of the solid electrodes, concentration dependences of the peak c1 were measured in ammonium formate, phosphate buffer pH 6.9 at the m-AgSAE, or in sodium acetate buffer pH 5.0 at the bPGE. LOD of 5 & micro;M was achieved with all three electrodes in the buffer solutions. Using HMDE, the same LOD was attained also in cell culturing media without any sample pretreatment. Incorporation of 2 '-deoxyzebularine 5 '-triphosphate into DNA was studied using primer extension and terminal deoxynucleotidyl transferase tailing techniques. Incorporation of Zeb exhibited poor efficacy; nevertheless, products of TdT tailing with deoxyZeb were successfully detected by ex situ CV. Interestingly, another epigenetic modulator, 5-azacytidine showed facile incorporation in both approaches. These observations were in accord with the presumably different mechanism of action of the two epigenetic drugs in cells. We demonstrate usefulness of a simple electrochemical method to assess different facileness of polymerase incorporation of two epigenetic drugs, cytidine analogues Zeb and 5-aC, by DNA polymerases.
ER  -

DAŇHEL, Aleš; Peter ŠEBEST; Daniel DOBROVODSKÝ; Marek ZELINKA; Pavlina HAVRANOVA; Luděk HAVRAN; Miloslava FOJTOVÁ a Miroslav FOJTA. Electrochemistry of epigenetic modulator zebularine: Voltammetric determination in cell culturing media and monitoring of polymerase incorporation into DNA. \textit{ELECTROCHIMICA ACTA}. OXFORD: PERGAMON-ELSEVIER SCIENCE LTD, 2026, roč.~563, s.~1-10. ISSN~0013-4686. Dostupné z: https://doi.org/10.1016/j.electacta.2026.148707.

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