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

J 2019

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

HERMANOVA, M.; P. ORSAG; J. BALINTOVA; M. HOCEK; Miroslav FOJTA et. al.

Základní údaje

Originální název

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

Autoři

HERMANOVA, M. (203 Česká republika); P. ORSAG (203 Česká republika); J. BALINTOVA (203 Česká republika); M. HOCEK (203 Česká republika) a Miroslav FOJTA (203 Česká republika, garant, domácí)

Vydání

Analytica Chimica Acta, Amsterdam, Elsevier Science publishers, 2019, 0003-2670

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.977

Kód RIV

RIV/00216224:14740/19:00108148

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1016/j.aca.2018.10.053

UT WoS

000455933400014

EID Scopus

2-s2.0-85055755902

Klíčová slova anglicky

DNA modification; Redox coding; DNA-protein interactions; Immunoprecipitation; Voltammetry

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 31. 3. 2020 21:32, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Návaznosti

GBP206/12/G151, projekt VaV

Název: Centrum nových přístupů k bioanalýze a molekulární diagnostice

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

Citovat

HERMANOVA, M.; P. ORSAG; J. BALINTOVA; M. HOCEK a Miroslav FOJTA. Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions. Analytica Chimica Acta. Amsterdam: Elsevier Science publishers, 2019, roč. 1050, MAR, s. 123-131. ISSN 0003-2670. Dostupné z: https://dx.doi.org/10.1016/j.aca.2018.10.053.

@article{1621364,
   author = {Hermanova, M. and Orsag, P. and Balintova, J. and Hocek, M. and Fojta, Miroslav},
   article_location = {Amsterdam},
   article_number = {MAR},
   doi = {http://dx.doi.org/10.1016/j.aca.2018.10.053},
   keywords = {DNA modification; Redox coding; DNA-protein interactions; Immunoprecipitation; Voltammetry},
   language = {eng},
   issn = {0003-2670},
   journal = {Analytica Chimica Acta},
   title = {Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions},
   url = {https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub},
   volume = {1050},
   year = {2019}
}

TY  - JOUR
ID  - 1621364
AU  - Hermanova, M. - Orsag, P. - Balintova, J. - Hocek, M. - Fojta, Miroslav
PY  - 2019
TI  - Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions
JF  - Analytica Chimica Acta
VL  - 1050
IS  - MAR
SP  - 123-131
EP  - 123-131
PB  - Elsevier Science publishers
SN  - 00032670
KW  - DNA modification
KW  - Redox coding
KW  - DNA-protein interactions
KW  - Immunoprecipitation
KW  - Voltammetry
UR  - https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub
L2  - https://www.sciencedirect.com/science/article/pii/S0003267018312959?via%3Dihub
N2  - 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.
ER  -

HERMANOVA, M.; P. ORSAG; J. BALINTOVA; M. HOCEK a Miroslav FOJTA. Dual redox labeling of DNA as a tool for electrochemical detection of p53 protein-DNA interactions. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier Science publishers, 2019, roč.~1050, MAR, s.~123-131. ISSN~0003-2670. Dostupné z: https://dx.doi.org/10.1016/j.aca.2018.10.053.

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