High-throughput Assays for Superoxide and Hydrogen Peroxide

@article{Zielonka2014HighthroughputAF,
  title={High-throughput Assays for Superoxide and Hydrogen Peroxide},
  author={Jacek Zielonka and Gang Cheng and Monika Zielonka and Thota Ganesh and Aiming Sun and Joy Joseph and Radosław Michalski and William James O'brien and John David Lambeth and Balaraman Kalyanaraman},
  journal={The Journal of Biological Chemistry},
  year={2014},
  volume={289},
  pages={16176 - 16189}
}
Background: NADPH oxidases (Nox) are involved in pathogenesis of inflammatory and fibrotic diseases. Results: High-throughput screening methodology is developed for discovery and determination of Nox inhibitors and mechanism. Conclusion: Rapid and specific monitoring of superoxide and hydrogen peroxide is possible, enabling high-throughput screening of Nox isoform-specific inhibitors. Significance: The proposed strategy will enable more rigorous research on the chemical biology of Nox enzymes… 
View on ASBMB
jbc.org
54 Citations
Highly Influential Citations
3
Background Citations
19
Methods Citations
11
Results Citations
1

Tables from this paper

54 Citations

Mitigation of NADPH Oxidase 2 Activity as a Strategy to Inhibit Peroxynitrite Formation*

It is concluded that the newly developed high throughput screening/reactive oxygen species assays could also be used to identify potential inhibitors of ONOO− formed from Nox2-derived O2̇̄ and nitric oxide synthase-derived nitricoxide.

Recent developments in detection of superoxide radical anion and hydrogen peroxide: Opportunities, challenges, and implications in redox signaling.

Pharmacological characterization of the seven human NOX isoforms and their inhibitors

Detection and Characterization of Reactive Oxygen and Nitrogen Species in Biological Systems by Monitoring Species-Specific Products.

Formation and detection of species-specific products from the probes enables accurate characterization of the oxidative environment in cells and provides a rigorous method for detection of those species in biological systems.

Small-molecule luminescent probes for the detection of cellular oxidizing and nitrating species.

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

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

The development of new assays for measuring the activity of NADPH oxidases enabling the high-throughput screening for NAD PH oxidase inhibitors is described.

Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection

It is demonstrated that the co-application of the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence without affecting the background, highlighting a novel feature of compounds containing the peptides boronics acid as powerful enhancers for the detection of superoxide.

NADPH oxidases as drug targets and biomarkers in neurodegenerative diseases: What is the evidence?

Isoform-selective NADPH oxidase inhibitor panel for pharmacological target validation.

...

References

SHOWING 1-10 OF 61 REFERENCES

Global Profiling of Reactive Oxygen and Nitrogen Species in Biological Systems

This global profiling approach, simultaneous detection of multiple ROS/RNS products of fluorescent probes, developed in this study will be useful in unraveling the complex role of ROS/ RNS in redox regulation, cell signaling, and cellular oxidative processes and in high-throughput screening of anti-inflammatory antioxidants.

Small-molecule NOX inhibitors: ROS-generating NADPH oxidases as therapeutic targets.

It is concluded that NOX inhibition is a pertinent and promising novel pharmacologic concept, but that major efforts will be necessary to develop specific NOX inhibitors suited for clinical application.

NOX enzymes as novel targets for drug development

The time is here for vigorous efforts to develop clinically useful inhibitors of the NOX enzymes, as most (though not all) NOX-related diseases appear to be mediated by a single member of theNOX family, agents with isoform specificity will be preferred, although broadly active NOX inhibitors may be useful in some settings.

A novel and specific NADPH oxidase-1 (Nox1) small-molecule inhibitor blocks the formation of functional invadopodia in human colon cancer cells.

The results elucidate the relevance of Nox1-dependent ROS generation in mechanisms of cancer invasion and define ML171 as a useful Nox 1 chemical probe and potential therapeutic agent for inhibition of cancer cell invasion.

Oxidases and peroxidases in cardiovascular and lung disease: new concepts in reactive oxygen species signaling.

First in class, potent, and orally bioavailable NADPH oxidase isoform 4 (Nox4) inhibitors for the treatment of idiopathic pulmonary fibrosis.

The design, synthesis, and optimization of first-in-class series of inhibitors of NADPH oxidase isoform 4 (Nox4), an enzyme implicated in several pathologies, allowing further clinical trials for the potential treatment of fibrotic diseases, cancers, and cardiovascular and metabolic diseases.

Promyelocytic HL60 cells express NADPH oxidase and are excellent targets in a rapid spectrophotometric microplate assay for extracellular superoxide.

It is shown that nondifferentiated HL60 cells contain NADPH oxidase and can be used as a model for the assessment of oxidant as well as antioxidant compounds and established a highly specific, reproducible and time-saving superoxide microplate assay as a valuable tool for the rapid screening of compounds for oxidative and antioxidants activity.

Oxidative chemistry of fluorescent dyes: implications in the detection of reactive oxygen and nitrogen species.

It is concluded that flurorescence microscopy or related techniques are not sufficient to measure the superoxide-specific hydroxylated products and tests using fluorescent-based boronate probes will be more reliable for peroxynitrite determination.

NOX4 activity is determined by mRNA levels and reveals a unique pattern of ROS generation.

NOX4 probably generates O(2)(-) within an intracellular compartment that is accessible to NBT (Nitro Blue Tetrazolium), but not to DHE or ACP, whereas a robust signal was observed with NBT.

Nox isoforms in vascular pathophysiology: insights from transgenic and knockout mouse models

The literature on studies utilising genetically-modified mouse strains to investigate the roles of NADPH oxidases in experimental models of vascular disease is reviewed to provide direction for future programs aimed at developing selective inhibitors of these enzymes as novel therapeutics in cardiovascular disease.
...