Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F17%3A00476545" target="_blank" >RIV/68081707:_____/17:00476545 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.bioelechem.2016.12.003" target="_blank" >http://dx.doi.org/10.1016/j.bioelechem.2016.12.003</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bioelechem.2016.12.003" target="_blank" >10.1016/j.bioelechem.2016.12.003</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer

Popis výsledku v původním jazyce

Specific DNA-protein interactions are vital for cellular life maintenance processes, such as transcriptional regulation, chromosome maintenance, replication and DNA repair, and their monitoring gives valuable information on molecular-level organization of those processes. Here, we propose a new method of label-free electrochemical sensing of sequence specific binding between the lysozyme protein and a single stranded DNA aptamer specific for lysozyme (DNA(apta)) that exploits the constant current chronopotentiometric stripping (CPS) analysis at modified mercury electrodes. Specific lysozyme-DNA(apta) binding Was distinguished from nonspecific lysozyme-DNA interactions at thioglycolic acid-modified mercury electrodes, but not at the dithiothreitol-modified or bare mercury electrodes. Stability of the surface-attached lysozyme-DNA(apta), layer depended on the stripping current (I-str) intensity, suggesting that the integrity of the layer critically depends on the time of its exposure to negative potentials. Stabilities of different lysozyme-DNA complexes at the negatively polarized electrode surface were tested, and it was shown that structural transitions of the specific lysozyme-DNA(apta) complexes occur in the I-str ranges different from those observed for assemblies of lysozyme with DNA sequences capable of only nonspecific lysozyme-DNA interactions. Thus, the CPS allows distinct discrimination between specific and non-specific protein-DNA binding and provides valuable information on stability of the nucleic acid-protein interactions at the polarized interfaces. (C) 2016 Elsevier B.V. All rights reserved.

Název v anglickém jazyce

Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer

Popis výsledku anglicky

Specific DNA-protein interactions are vital for cellular life maintenance processes, such as transcriptional regulation, chromosome maintenance, replication and DNA repair, and their monitoring gives valuable information on molecular-level organization of those processes. Here, we propose a new method of label-free electrochemical sensing of sequence specific binding between the lysozyme protein and a single stranded DNA aptamer specific for lysozyme (DNA(apta)) that exploits the constant current chronopotentiometric stripping (CPS) analysis at modified mercury electrodes. Specific lysozyme-DNA(apta) binding Was distinguished from nonspecific lysozyme-DNA interactions at thioglycolic acid-modified mercury electrodes, but not at the dithiothreitol-modified or bare mercury electrodes. Stability of the surface-attached lysozyme-DNA(apta), layer depended on the stripping current (I-str) intensity, suggesting that the integrity of the layer critically depends on the time of its exposure to negative potentials. Stabilities of different lysozyme-DNA complexes at the negatively polarized electrode surface were tested, and it was shown that structural transitions of the specific lysozyme-DNA(apta) complexes occur in the I-str ranges different from those observed for assemblies of lysozyme with DNA sequences capable of only nonspecific lysozyme-DNA interactions. Thus, the CPS allows distinct discrimination between specific and non-specific protein-DNA binding and provides valuable information on stability of the nucleic acid-protein interactions at the polarized interfaces. (C) 2016 Elsevier B.V. All rights reserved.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA13-00956S" target="_blank" >GA13-00956S: Úloha proteinů rodiny Anterior gradient při vzniku a vývoji nádorových onemocnění</a><br>

Návaznosti

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

Rok uplatnění

2017

Kód důvěrnosti údajů

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

Název periodika

Bioelectrochemistry

ISSN

1567-5394

e-ISSN

—

Svazek periodika

114

Číslo periodika v rámci svazku

APR2017

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

6

Strana od-do

42-47

Kód UT WoS článku

000394073500006

EID výsledku v databázi Scopus

—