Fluorescent probes for monitoring myeloperoxidase-derived hypochlorous acid: a comparative study

Karolina Pierzchała; Marlena Pięta; Monika Rola; Małgorzata Świerczyńska; Angelika Artelska; Karolina Dębowska; Radosław Podsiadły; Jakub Pięta; Jacek Zielonka; Adam Sikora; Andrzej Marcinek; Radosław Michalski

doi:10.1038/s41598-022-13317-8

Scientific Reports (Jun 2022)

Fluorescent probes for monitoring myeloperoxidase-derived hypochlorous acid: a comparative study

Karolina Pierzchała,
Marlena Pięta,
Monika Rola,
Małgorzata Świerczyńska,
Angelika Artelska,
Karolina Dębowska,
Radosław Podsiadły,
Jakub Pięta,
Jacek Zielonka,
Adam Sikora,
Andrzej Marcinek,
Radosław Michalski

Affiliations

Karolina Pierzchała: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Marlena Pięta: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Monika Rola: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Małgorzata Świerczyńska: Department of Chemistry, Institute of Polymer and Dye Technology, Lodz University of Technology
Angelika Artelska: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Karolina Dębowska: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Radosław Podsiadły: Department of Chemistry, Institute of Polymer and Dye Technology, Lodz University of Technology
Jakub Pięta: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Jacek Zielonka: Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin
Adam Sikora: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Andrzej Marcinek: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology
Radosław Michalski: Department of Chemistry, Institute of Applied Radiation Chemistry, Lodz University of Technology

DOI: https://doi.org/10.1038/s41598-022-13317-8
Journal volume & issue: Vol. 12, no. 1
pp. 1 – 14

Abstract

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Abstract MPO-derived oxidants including HOCl contribute to tissue damage and the initiation and propagation of inflammatory diseases. The search for small molecule inhibitors of myeloperoxidase, as molecular tools and potential drugs, requires the application of high throughput screening assays based on monitoring the activity of myeloperoxidase. In this study, we have compared three classes of fluorescent probes for monitoring myeloperoxidase-derived hypochlorous acid, including boronate-, aminophenyl- and thiol-based fluorogenic probes and we show that all three classes of probes are suitable for this purpose. However, probes based on the coumarin fluorophore turned out to be not reliable indicators of the inhibitors’ potency. We have also determined the rate constants of the reaction between HOCl and the probes and they are equal to 1.8 × 104 M−1s−1 for coumarin boronic acid (CBA), 1.1 × 104 M−1s−1 for fluorescein based boronic acid (FLBA), 3.1 × 104 M−1s−1 for 7-(p-aminophenyl)-coumarin (APC), 1.6 × 104 M−1s−1 for 3’-(p-aminophenyl)-fluorescein (APF), and 1 × 107 M−1s−1 for 4-thiomorpholino-7-nitrobenz-2-oxa-1,3-diazole (NBD-TM). The high reaction rate constant of NBD-TM with HOCl makes this probe the most reliable tool to monitor HOCl formation in the presence of compounds showing HOCl-scavenging activity.

Published in Scientific Reports

ISSN: 2045-2322 (Online)
Publisher: Nature Portfolio
Country of publisher: United Kingdom
LCC subjects: Medicine; Science
Website: https://www.nature.com/srep/

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