Detailed Information on Publication Record
2014
Electrochemical detection of DNA binding by tumor suppressor p53 protein using osmium-labeled oligonucleotide probes and catalytic hydrogen evolution at the mercury electrode
NĚMCOVÁ, Kateřina, Peter ŠEBEST, Luděk HAVRAN, Petr ORSÁG, Miroslav FOJTA et. al.Basic information
Original name
Electrochemical detection of DNA binding by tumor suppressor p53 protein using osmium-labeled oligonucleotide probes and catalytic hydrogen evolution at the mercury electrode
Authors
NĚMCOVÁ, Kateřina (203 Czech Republic), Peter ŠEBEST (203 Czech Republic), Luděk HAVRAN (203 Czech Republic), Petr ORSÁG (203 Czech Republic, belonging to the institution), Miroslav FOJTA (203 Czech Republic, guarantor, belonging to the institution) and Hana PIVOŇKOVÁ (203 Czech Republic)
Edition
Analytical and Bioanalytical chemistry, HEIDELBERG, SPRINGER HEIDELBERG, 2014, 1618-2642
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10405 Electrochemistry
Country of publisher
Germany
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 3.436
RIV identification code
RIV/00216224:14740/14:00079183
Organization unit
Central European Institute of Technology
UT WoS
000341834500011
Keywords in English
Electrochemical analysis; Labeled probes; Osmium complex; Tumor suppressor protein p53; DNA-protein interaction; Immunoprecipitation; Competition assay; Magnetic beads; Mercury electrode
Tags
Tags
International impact, Reviewed
Změněno: 23/2/2015 13:18, Martina Prášilová
Abstract
V originále
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. Versatility of the electrochemical MBIP technique and its general applicability in studies of any DNA-binding protein is discussed.