Počet záznamů: 1
Electrochemical detection of DNA binding by tumor suppressor p53 protein using osmium-labeled oligonucleotide probes and catalytic hydrogen evolution at the mercury electrode
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SYSNO ASEP 0435469 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Electrochemical detection of DNA binding by tumor suppressor p53 protein using osmium-labeled oligonucleotide probes and catalytic hydrogen evolution at the mercury electrode Tvůrce(i) Němcová, Kateřina (BFU-R)
Šebest, Peter (BFU-R) ORCID, RID
Havran, Luděk (BFU-R) RID, ORCID
Orság, Petr (BFU-R) ORCID
Fojta, Miroslav (BFU-R) RID, ORCID
Pivoňková, Hana (BFU-R) ORCID, RIDCelkový počet autorů 6 Zdroj.dok. Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 406, č. 24 (2014), s. 5843-5852Poč.str. 10 s. Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. DE - Německo Klíč. slova Electrochemical analysis ; Labeled probes ; Osmium complex Vědní obor RIV BO - Biofyzika CEP GAP301/11/2076 GA ČR - Grantová agentura ČR IAA400040901 GA AV ČR - Akademie věd Institucionální podpora BFU-R - RVO:68081707 UT WOS 000341834500011 DOI https://doi.org/10.1007/s00216-014-7996-0 Anotace In this paper, we present an electrochemical DNA-protein interaction assay based on a combination of protein-specific immunoprecipitation at magnetic beads (MBIP) with application of oligonucleotide (ON) probes labeled with an electroactive oxoosmium complex (Os,bipy). We show that double-stranded ONs bearing a dT(20) tail labeled with Os,bipy are specifically recognized by the tumor suppressor p53 protein according to the presence or absence of a specific binding site (p53CON) in the double-stranded segment. We demonstrate the applicability of the Os,bipy-labeled probes in titration as well as competition MBIP assays to evaluate p53 relative affinity to various sequence-specific or structurally distinct unlabeled DNA substrates upon modulation of the p53-DNA binding by monoclonal antibodies used for the immunoprecipitation. To detect the p53-bound osmium-labeled probes, we took advantage of a catalytic peak yielded by Os,bipy-modified DNA at the mercury-based electrodes, allowing facile determination of subnanogram quantities of the labeled oligonucleotides. Pracoviště Biofyzikální ústav Kontakt Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Rok sběru 2015
Počet záznamů: 1