Počet záznamů: 1
Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer
- 1. 0476545 - BFU-R 2018 RIV CH eng J - Článek v odborném periodiku
Ostatná, Veronika - Vargová, Veronika - Kekedy-Nagy, L. - Černocká, Hana - Ferapontova, E.E.
Chronopotentiometric sensing of specific interactions between lysozyme and the DNA aptamer.
Bioelectrochemistry. Roč. 114, APR2017 (2017), s. 42-47. ISSN 1567-5394
Grant CEP: GA ČR GA13-00956S
Institucionální podpora: RVO:68081707
Klíčová slova: self-assembled monolayers * protein interactions * lysozyme
Kód oboru RIV: CE - Biochemie
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 3.789, rok: 2017
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.
Trvalý link: http://hdl.handle.net/11104/0273024