Recent Developments in the Probes and Assays for Measurement of the Activity of NADPH Oxidases

Zielonka, Jacek; Hardy, Micaël; Michalski, Radosław; Sikora, Adam; Zielonka, Monika; Cheng, Gang; Ouari, Olivier; Podsiadly, Radosław; Kalyanaraman, Balaraman

doi:10.1007/s12013-017-0813-6

Cited by 23 publications

(23 citation statements)

References 106 publications

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“…•− and H 2 O 2 by the probes themselves (Zielonka et al, 2013b(Zielonka et al, , 2017aKalyanaraman et al, 2016). Boronate redox probes have emerged as a new tool to study the activity and role of NADPH oxidases in cellular redox signaling and immune response (Dickinson et al, 2011a;Brewer et al, 2015;Zielonka et al, 2017a).…”

Section: Boronate-based Redox Probes In the Studies Of Nadph Oxidasesmentioning

confidence: 99%

“…In addition to mitochondrial electron transport chain, NADPH oxidases (or Nox enzymes) are regarded as the major source of cellular

and H 2 O 2 . While an array of probes and assays have been developed over the last 30 years, in order to monitor Nox activity, most probes suffer from serious limitations, including the potential to generate

and H 2 O 2 by the probes themselves (Zielonka et al, 2013b , 2017a ; Kalyanaraman et al, 2016 ). Boronate redox probes have emerged as a new tool to study the activity and role of NADPH oxidases in cellular redox signaling and immune response (Dickinson et al, 2011a ; Brewer et al, 2015 ; Zielonka et al, 2017a ).…”

Section: Boronate-based Redox Probes In the Studies Of Nadph Oxidasesmentioning

confidence: 99%

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Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology

et al. 2020

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Boronate-based molecular probes are emerging as one of the most effective tools for detection and quantitation of peroxynitrite and hydroperoxides. This review discusses the chemical reactivity of boronate compounds in the context of their use for detection of biological oxidants, and presents examples of the practical use of those probes in selected chemical, enzymatic, and biological systems. The particular reactivity of boronates toward nucleophilic oxidants makes them a distinct class of probes for redox biology studies. We focus on the recent progress in the design and application of boronate-based probes in redox studies and perspectives for further developments.

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Section: Boronate-based Redox Probes In the Studies Of Nadph Oxidasesmentioning

confidence: 99%

“…In addition to mitochondrial electron transport chain, NADPH oxidases (or Nox enzymes) are regarded as the major source of cellular

Section: Boronate-based Redox Probes In the Studies Of Nadph Oxidasesmentioning

confidence: 99%

Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology

et al. 2020

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“…However, the ability of those treatments to induce O2 •generation in the used cell model has not been shown. Numerous reports demonstrate the utility of HE, MitoSOX Red, and hydropropidine, when coupled with HPLC-based analyses, to detect O2 •in different cellular models, as reviewed elsewhere (20,21,63). In those reports, 2-OH-E + , 2-OH-Mito-E + , and 2-OH-Pr ++ were used as specific marker products for O2 •-.…”

Section: -Hydroxyethidium As a Specific Marker For O2 •-mentioning

confidence: 99%

Tracking isotopically labeled oxidants using boronate-based redox probes

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Kalyanaraman

et al. 2020

Journal of Biological Chemistry

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Reactive oxygen and nitrogen species have been implicated in many biological processes and diseases, including immune responses, cardiovascular dysfunction, neurodegeneration, and cancer. These chemical species are short-lived in biological settings, and detecting them in these conditions and diseases requires the use of molecular probes that form stable, easily detectable, products. The chemical mechanisms and limitations of many of the currently used probes are not well-understood, hampering their effective applications. Boronates have emerged as a class of probes for the detection of nucleophilic two-electron oxidants. Here, we report the results of an oxygen-18–labeling MS study to identify the origin of oxygen atoms in the oxidation products of phenylboronate targeted to mitochondria. We demonstrate that boronate oxidation by hydrogen peroxide, peroxymonocarbonate, hypochlorite, or peroxynitrite involves the incorporation of oxygen atoms from these oxidants. We therefore conclude that boronates can be used as probes to track isotopically labeled oxidants. This suggests that the detection of specific products formed from these redox probes could enable precise identification of oxidants formed in biological systems. We discuss the implications of these results for understanding the mechanism of conversion of the boronate-based redox probes to oxidant-specific products.

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“…A reaction between O 2 •– and hydroethidine (HE) results in the formation of a very specific product, 2-hydroxyethidium (2-OH-E + ); this product is not formed from the reaction between HE and other biologically relevant oxidants such as H 2 O 2 , singlet oxygen, lipid hydroperoxides, peroxynitrite, HOCl, and • NO 2 [14] , [15] . This marker product derived from the O 2 •– and HE reaction (2-OH-E + ) can be unambiguously detected by rapid high-performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC-MS) methods [16] . However, numerous publications using HE-derived fluorescence still posit that ethidium (E + ) is the product of the O 2 •– reaction with HE, whereas it has been clearly established by us and others that E + is not the product of the reaction between O 2 •– and HE ( Fig.…”

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Teaching the basics of reactive oxygen species and their relevance to cancer biology: Mitochondrial reactive oxygen species detection, redox signaling, and targeted therapies

et al. 2018

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Reactive oxygen species (ROS) have been implicated in tumorigenesis (tumor initiation, tumor progression, and metastasis). Of the many cellular sources of ROS generation, the mitochondria and the NADPH oxidase family of enzymes are possibly the most prevalent intracellular sources. In this article, we discuss the methodologies to detect mitochondria-derived superoxide and hydrogen peroxide using conventional probes as well as newly developed assays and probes, and the necessity of characterizing the diagnostic marker products with HPLC and LC-MS in order to rigorously identify the oxidizing species. The redox signaling roles of mitochondrial ROS, mitochondrial thiol peroxidases, and transcription factors in response to mitochondria-targeted drugs are highlighted. ROS generation and ROS detoxification in drug-resistant cancer cells and the relationship to metabolic reprogramming are discussed. Understanding the subtle role of ROS in redox signaling and in tumor proliferation, progression, and metastasis as well as the molecular and cellular mechanisms (e.g., autophagy) could help in the development of combination therapies. The paradoxical aspects of antioxidants in cancer treatment are highlighted in relation to the ROS mechanisms in normal and cancer cells. Finally, the potential uses of newly synthesized exomarker probes for in vivo superoxide and hydrogen peroxide detection and the low-temperature electron paramagnetic resonance technique for monitoring oxidant production in tumor tissues are discussed.

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Recent Developments in the Probes and Assays for Measurement of the Activity of NADPH Oxidases

Cited by 23 publications

References 106 publications

Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology

Boronate-Based Probes for Biological Oxidants: A Novel Class of Molecular Tools for Redox Biology

Tracking isotopically labeled oxidants using boronate-based redox probes

Teaching the basics of reactive oxygen species and their relevance to cancer biology: Mitochondrial reactive oxygen species detection, redox signaling, and targeted therapies

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