Cited by 180Redox biology and oxidative stressNeutrophil, Myeloperoxidase and Oxidative MechanismsNitric Oxide and Endothelin Effects

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Detection of superoxide anion and hydrogen peroxide production by cellular NADPH oxidases

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Nauseef, William M.·2014·Biochimica et Biophysica Acta (BBA) - General Subjects

DOI 10.1016/j.bbagen.2013.04.040PMC PMC3770773

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Abstract

Background The recent recognition that isoforms of the cellular NADPH-dependent oxidases, collectively known as the NOX protein family, participate in a wide range of physiologic and pathophysiologic processes in both the animal and plant kingdoms has stimulated interest in the identification, localization, and quantitation of their products in biological settings. Although several tools for measuring oxidants released extracellularly are available, the specificity and selectivity of the methods for reliable analysis of intracellular oxidants have not matched the enthusiasm for studying NOX proteins.Scope of review Focusing exclusively on superoxide anion and hydrogen peroxide produced by NOX proteins, this review describes the ideal probe for analysis of O2(-) and H2O2 generated extracellularly and intracellularly by NOX proteins. An overview of the components, organization, and topology of NOX proteins provides a rationale for applying specific probes for use and a context in which to interpret results and thereby construct plausible models linking NOX-derived oxidants to biological responses. The merits and shortcomings of methods currently in use to assess NOX activity are highlighted, and those assays that provide quantitation of superoxide or H2O2 are contrasted with those intended to examine spatial and temporal aspects of NOX activity.Major conclusions Although interest in measuring the extracellular and intracellular products of the NOX protein family is great, robust analytical probes are limited.General significance The widespread involvement of NOX proteins in many biological processes requires rigorous approaches to the detection, localization, and quantitation of the oxidants produced. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.

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Authors: Nauseef, W. M.
Affiliations: University of Iowa (US)
Corresponding author: Nauseef, William M.
ORCID: Nauseef, William M.
Venue: Biochimica et Biophysica Acta (BBA) - General Subjects
Publisher: Elsevier BV
Published: 2014-02
Volume: 1840
Issue: 2
Pages: 757-767
ISSN: 0304-4165 (ISSN-L)
DOI: 10.1016/j.bbagen.2013.04.040
PMC: PMC3770773
OpenAlex: W2085864038
Cited by: 180
References: 171
Citations by year: 2026: 11 · 2025: 9 · 2024: 13 · 2023: 12 · 2022: 17 · 2021: 5 · 2020: 16 · 2019: 14 · 2018: 14 · 2017: 19 · 2016: 17 · 2015: 17 · 2014: 13
FWCI: 1.53
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations: Biochimica et Biophysica Acta (BBA) - General Subjects (publishedVersion)
PubMed (publishedVersion)
PubMed Central (acceptedVersion)
Funders: National Institute of Health (AI70958, AI044642)
Veterans Administration
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
Keywords: NADPH oxidase, superoxide, hydrogen peroxide, reactive oxygen species, fluorescent probes
MeSH terms: Animals, Humans, Hydrogen Peroxide, Superoxides, NADPH Oxidases

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Detection of superoxide anion and hydrogen peroxide production by cellular NADPH oxidases

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