Interfacial properties of p53-DNA complexes containing various recognition elements

Methods which can distinguish between specific and non-specific protein interactions leading to the identification of hubs and nodes are still desired. This work shows utilization of chronopotentiometric stripping analysis in combination with a mercury electrode in the study of protein-DNA interacti...

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Published in:Journal of electroanalytical chemistry (Lausanne, Switzerland) Switzerland), 2019-09, Vol.848, p.113300, Article 113300
Main Authors: Černocká, Hana, Fojt, Lukáš, Adámik, Matej, Brázdová, Marie, Paleček, Emil, Ostatná, Veronika
Format: Article
Language:English
Subjects:
Apoptosis
Binding sites
Chemical reduction
Constant current chronopotentiometry
Deoxyribonucleic acid
Deoxyribonucleic acid-protein binding
Disintegration
DNA
Electric fields
Electrochemical sensing
Electrodes
Gels
Interfacial properties
Mercury containing electrodes
Polyacrylamide
Proteins
Recognition
Stripping
Tumor suppressor protein p53
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Summary:Methods which can distinguish between specific and non-specific protein interactions leading to the identification of hubs and nodes are still desired. This work shows utilization of chronopotentiometric stripping analysis in combination with a mercury electrode in the study of protein-DNA interactions at thiol-modified electrodes. The complex of tumor suppressor p53 core domain (p53CD) and DNA undergoes disintegration due to the effect of the electric field, accompanied by a remarkable increase in the electrocatalytic reduction signal. By adjusting stripping current intensities and temperature, the transition between intact and disintegrated complex reflected differences in the stabilities of sequence-specific complexes with different recognition elements. Higher stabilities of p53-DNA complexes were observed for DNA binding sites connected with cell-cycle arrest and p53 negative autoregulation, than those for DNA associated with cell apoptosis, in good concordance with electrophoretic mobility shift assay in polyacrylamide gels. These data highlight the utility of this method for studying the dynamics of surface-attached protein-DNA complexes. •DNA binding to oncoprotein p53 was detected by constant current chronopotentiometry.•Surface-attached DNA-p53 complexes on mercury disintegrate at negative potentials.•Complexes of p53 core domain with various sequence-specific DNA were discriminated.
ISSN:1572-6657
1873-2569
DOI:10.1016/j.jelechem.2019.113300