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Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer
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SYSNO ASEP 0476545 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer Author(s) Ostatná, Veronika (BFU-R) RID, ORCID
Vargová, Veronika (BFU-R)
Kekedy-Nagy, L. (DK)
Černocká, Hana (BFU-R) RID, ORCID
Ferapontova, E.E. (DK)Number of authors 5 Source Title Bioelectrochemistry. - : Elsevier - ISSN 1567-5394
Roč. 114, APR2017 (2017), s. 42-47Number of pages 6 s. Publication form Print - P Language eng - English Country CH - Switzerland Keywords self-assembled monolayers ; protein interactions ; lysozyme Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology R&D Projects GA13-00956S GA ČR - Czech Science Foundation (CSF) Institutional support BFU-R - RVO:68081707 UT WOS 000394073500006 DOI 10.1016/j.bioelechem.2016.12.003 Annotation 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. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2018
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