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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F19%3A00108148" target="_blank" >RIV/00216224:14740/19:00108148 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68081707:_____/19:00501012 RIV/61388963:_____/19:00501012 RIV/00216208:11310/19:10394770

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.aca.2018.10.053" target="_blank" >10.1016/j.aca.2018.10.053</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Analytica Chimica Acta

ISSN

0003-2670

e-ISSN

—

Svazek periodika

1050

Číslo periodika v rámci svazku

MAR

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

9

Strana od-do

123-131

Kód UT WoS článku

000455933400014

EID výsledku v databázi Scopus

2-s2.0-85055755902