Abstract
The development of boronic probes enabled reliable detection and quantitative analysis of hydrogen peroxide , other nucleophilic hydroperoxides, hypochlorite , and peroxynitrite . The major product, in which boronate moiety of the probe is replaced by the hydroxyl group, is, however, common for all those oxidants. Here, we describe how ortho-isomer of mitochondria-targeted phenylboronic acid can be used to detect and differentiate peroxynitrite-dependent and independent probe oxidation. This method highlights detection and quantification of both the major, phenolic product and the minor, peroxynitrite-specific cyclic and nitrated products of probe oxidation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Lippert AR, Van de Bittner GC, Chang CJ (2011) Boronate oxidation as a bioorthogonal reaction approach for studying the chemistry of hydrogen peroxide in living systems. Acc Chem Res 44:793–804
Zielonka J, Sikora A, Hardy M, Joseph J, Dranka BP, Kalyanaraman B (2012) Boronate probes as diagnostic tools for real time monitoring of peroxynitrite and hydroperoxides. Chem Res Toxicol 25:1793–1799
Sikora A, Zielonka J, Debowska K, Michalski R, Smulik-Izydorczyk R, Pieta J, Podsiadly R, Artelska A, Pierzchala K, Kalyanaraman B (2020) Boronate-based probes for biological oxidants: A novel class of molecular tools for redox biology. Front Chem 8:580899
Dickinson BC, Chang CJ (2008) A targetable fluorescent probe for imaging hydrogen peroxide in the mitochondria of living cells. J Am Chem Soc 130:9638–9639
Cocheme HM, Quin C, McQuaker SJ, Cabreiro F, Logan A, Prime TA, Abakumova I, Patel JV, Fearnley IM, James AM, Porteous CM, Smith RA, Saeed S, Carre JE, Singer M, Gems D, Hartley RC, Partridge L, Murphy MP (2011) Measurement of H2O2 within living Drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix. Cell Metab 13:340–350
Cocheme HM, Logan A, Prime TA, Abakumova I, Quin C, McQuaker SJ, Patel JV, Fearnley IM, James AM, Porteous CM, Smith RA, Hartley RC, Partridge L, Murphy MP (2012) Using the mitochondria-targeted ratiometric mass spectrometry probe MitoB to measure H2O2 in living Drosophila. Nat Protoc 7:946–958
Dickinson BC, Lin VS, Chang CJ (2013) Preparation and use of MitoPY1 for imaging hydrogen peroxide in mitochondria of live cells. Nat Protoc 8:1249–1259
Zielonka J, Joseph J, Sikora A, Hardy M, Ouari O, Vasquez-Vivar J, Cheng G, Lopez M, Kalyanaraman B (2017) Mitochondria-targeted triphenylphosphonium-based compounds: syntheses, mechanisms of action, and therapeutic and diagnostic applications. Chem Rev 117:10043–10120
Sikora A, Zielonka J, Lopez M, Joseph J, Kalyanaraman B (2009) Direct oxidation of boronates by peroxynitrite: mechanism and implications in fluorescence imaging of peroxynitrite. Free Radic Biol Med 47:1401–1407
Sikora A, Zielonka J, Lopez M, Dybala-Defratyka A, Joseph J, Marcinek A, Kalyanaraman B (2011) Reaction between peroxynitrite and boronates: EPR spin-trapping, HPLC analyses, and quantum mechanical study of the free radical pathway. Chem Res Toxicol 24:687–697
Wardman P (2007) Fluorescent and luminescent probes for measurement of oxidative and nitrosative species in cells and tissues: progress, pitfalls, and prospects. Free Radic Biol Med 43:995–1022
Kalyanaraman B (2011) Oxidative chemistry of fluorescent dyes: implications in the detection of reactive oxygen and nitrogen species. Biochem Soc Trans 39:1221–1225
Zielonka J, Kalyanaraman B (2018) Small-molecule luminescent probes for the detection of cellular oxidizing and nitrating species. Free Radic Biol Med 128:3–22
Zielonka J, Kalyanaraman B (2012) Methods of investigation of selected radical oxygen/nitrogen species in cell-free and cellular systems. In: Pantopoulos K, Schipper HM (eds) Principles of free radical biomedicine, vol I. Nova Science Publishers, New York, pp 201–264
Zielonka J, Sikora A, Joseph J, Kalyanaraman B (2010) Peroxynitrite is the major species formed from different flux ratios of co-generated nitric oxide and superoxide: direct reaction with boronate-based fluorescent probe. J Biol Chem 285:14210–14216
Michalski R, Zielonka J, Gapys E, Marcinek A, Joseph J, Kalyanaraman B (2014) Real-time measurements of amino acid and protein hydroperoxides using coumarin boronic acid. J Biol Chem 289:22536–22553
Truzzi DR, Augusto O (2017) Influence of CO2 on hydroperoxide metabolism. In: Vissers MC, Hampton M, Kettle AJ (eds) Hydrogen peroxide metabolism in health and disease. CRC Press, Boca Raton, FL, pp 81–99
Zielonka J, Zielonka M, Sikora A, Adamus J, Joseph J, Hardy M, Ouari O, Dranka BP, Kalyanaraman B (2012) Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem 287:2984–2995
Sikora A, Zielonka J, Adamus J, Debski D, Dybala-Defratyka A, Michalowski B, Joseph J, Hartley RC, Murphy MP, Kalyanaraman B (2013) Reaction between peroxynitrite and triphenylphosphonium-substituted arylboronic acid isomers: identification of diagnostic marker products and biological implications. Chem Res Toxicol 26:856–867
Zielonka J, Sikora A, Adamus J, Kalyanaraman B (2015) Detection and differentiation between peroxynitrite and hydroperoxides using mitochondria-targeted arylboronic acid. Methods Mol Biol 1264:171–181
Zielonka J, Zielonka M, VerPlank L, Cheng G, Hardy M, Ouari O, Ayhan MM, Podsiadły R, Sikora A, Lambeth JD, Kalyanaraman B (2016) Mitigation of NADPH oxidase 2 activity as a strategy to inhibit peroxynitrite formation. J Biol Chem 291:7029–7044
Rios N, Radi R, Kalyanaraman B, Zielonka J (2020) Tracking isotopically labeled oxidants using boronate-based redox probes. J Biol Chem 295:6665-6676
Zielonka J, Joseph J, Sikora A, Kalyanaraman B (2013) Real-time monitoring of reactive oxygen and nitrogen species in a multiwell plate using the diagnostic marker products of specific probes. Methods Enzymol 526:145–157
Hardy M, Zielonka J, Karoui H, Sikora A, Michalski R, Podsiadly R, Lopez M, Vasquez-Vivar J, Kalyanaraman B, Ouari O (2018) Detection and characterization of reactive oxygen and nitrogen species in biological systems by monitoring species-specific products. Antioxid Redox Signal 28:1416–1432
Cheng G, Zielonka M, Dranka B, Kumar SN, Myers CR, Bennett B, Garces AM, Dias Duarte Machado LG, Thiebaut D, Ouari O, Hardy M, Zielonka J, Kalyanaraman B (2018) Detection of mitochondria-generated reactive oxygen species in cells using multiple probes and methods: potentials, pitfalls, and the future. J Biol Chem 293:10363–10380
Morrison DE, Issa F, Bhadbhade M, Groebler L, Witting PK, Kassiou M, Rutledge PJ, Rendina LM (2010) Boronated phosphonium salts containing arylboronic acid, closo-carborane, or nido-carborane: synthesis, X-ray diffraction, in vitro cytotoxicity, and cellular uptake. J Biol Inorg Chem 15:1305–1318
Acknowledgments
This work was supported by NIH grant R01CA208648 (B.K.). JZ was supported in part by Institutional Research Grant IRG #16-183-31 from the American Cancer Society and the MCW Cancer Center. MH was supported, in part, by the French National Research Agency ANR-16-CE07-0023-01. AS was supported by Polish National Science Center within the SONATA BIS program (Grant Number 2015/18/E/ST4/00235). RP was supported by a grant from Polish National Science Centre (NCN) within the SONATA BIS 6 program (Grant no. 2016/22/E/ST4/00549).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Zielonka, J., Sikora, A., Podsiadly, R., Hardy, M., Kalyanaraman, B. (2021). Identification of Peroxynitrite by Profiling Oxidation and Nitration Products from Mitochondria-Targeted Arylboronic Acid . In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2275. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1262-0_20
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1262-0_20
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1261-3
Online ISBN: 978-1-0716-1262-0
eBook Packages: Springer Protocols