Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions

0501012 - BFÚ 2019 RIV NL eng J - Článek v odborném periodiku
Hermanová, Monika - Orság, Petr - Balintová, Jana - Hocek, Michal - Fojta, Miroslav
Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions.
Analytica Chimica Acta. Roč. 1050, Mar 7 (2019), s. 123-131. ISSN 0003-2670. E-ISSN 1873-4324
Grant CEP: GA ČR GBP206/12/G151
Grant ostatní: AV ČR(CZ) AP1501
Program: Akademická prémie - Praemium Academiae
Institucionální podpora: RVO:68081707 ; RVO:61388963
Klíčová slova: nucleoside triphosphates * enzymatic incorporation * binding * mutations
Obor OECD: Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis); Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis) (UOCHB-X)
Impakt faktor: 5.977, rok: 2019
Způsob publikování: Omezený přístup
https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub

We present a novel dual redox labeling approach enabling a facile relative evaluation of protein-DNA interactions based on immunoprecipitation at magnetic beads (MBIP) with subsequent electro-chemical detection. DNA probes labeled with two different electroactive markers, benzofurazane and nitrobenzene, which yield reduction peaks at distinct potentials, were synthesized using primer extension (PEX) reaction. We show that using the labeled DNA probes, specific and non-specific binding of the p53 protein can be distinguished in a simple competition binding experiment, as a strong preference of the p53 protein was observed towards DNA probes bearing a specific p53 binding site (p53CON), which is in agreement with known binding properties of the p53 protein. The p53 binding to the individual DNA probes can be modulated by specific monoclonal antibodies used for the immuno-precipitation. This approach can potentially be applied, after selection of appropriate DNA probes and monoclonal antibodies, for investigations of DNA-binding properties of other proteins and thus represents a versatile tool for studies of any DNA-binding proteins. (c) 2018 Elsevier B.V. All rights reserved.
Trvalý link: http://hdl.handle.net/11104/0293043