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Cysteamine assay for the evaluation of bioactive electrophiles
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SYSNO ASEP 0542079 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Cysteamine assay for the evaluation of bioactive electrophiles Author(s) Novák, D. (CZ)
Vrba, J. (CZ)
Zatloukalová, M. (CZ)
Roubalová, L. (CZ)
Stolarczyk, K. (CZ)
Dorčák, Vlastimil (BFU-R) RID, ORCID
Vacek, J. (CZ)Number of authors 7 Source Title Free Radical Biology and Medicine. - : Elsevier - ISSN 0891-5849
Roč. 164, FEB 20 2021 (2021), s. 381-389Number of pages 9 s. Publication form Print - P Language eng - English Country US - United States Keywords Thiol reactivity ; Cysteamine assay ; Electrochemistry ; Hydrophilic interaction chromatography ; Nrf2 pathway Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology R&D Projects GA19-09212S GA ČR - Czech Science Foundation (CSF) Method of publishing Limited access Institutional support BFU-R - RVO:68081707 UT WOS 000621327600002 EID SCOPUS 85099914970 DOI 10.1016/j.freeradbiomed.2021.01.007 Annotation Covalent modifications of thiol and amine groups may control the function of proteins involved in the regulatory and signaling pathways of the cell. In this study, we developed a simple cysteamine assay which can be used to study the reactivity of electrophilic compounds towards primary amine and thiol groups in an aqueous environment. The detection principle is based on the electrochemical, photometrical and mass spectrometric analyses of cysteamine (2-aminoethanethiol) as the molecular probe. This technique is useful for studying the reaction kinetics of electrophiles with thiol (SH) and amino (NH2) groups. The decrease in analytical responses of cysteamine was monitored to evaluate the reactivity of three electrophilic activators of the Nrf2 pathway, which mediates the cellular stress response. The SH-reactivity under cell-free conditions of the tested electrophiles decreased in the following order: 4-hydroxy-2-nonenal >= nitro-oleic acid > sulforaphane. However, as shown in RAW264.7 cells, the tested compounds activated Nrf2-dependent gene expression in the opposite order: sulforaphane > nitro-oleic acid >= 4-hydroxy-2-nonenal. Although other factors in addition to chemical reactivity play a role in biological systems, we conclude that this cysteamine assay is a useful tool for screening potentially bioactive electrophiles and for studying their reactivity at a molecular level. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2022 Electronic address https://www.sciencedirect.com/science/article/pii/S0891584921000137?via%3Dihub
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