Detection of reactive oxygen species derived from the family of NOX NADPH oxidases

NADPH oxidases (NOX) are superoxide anion radical (O2−•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O2−•. NOX have multiple...

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Published in:Free radical biology & medicine Vol. 53; no. 10; pp. 1903 - 1918
Main Authors: Maghzal, Ghassan J., Krause, Karl-Heinz, Stocker, Roland, Jaquet, Vincent
Format: Journal Article
Language:English
Published: United States Elsevier Inc 15.11.2012
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ISSN:0891-5849, 1873-4596, 1873-4596
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Abstract NADPH oxidases (NOX) are superoxide anion radical (O2−•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O2−•. NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O2−• or its direct dismutation product hydrogen peroxide (H2O2) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O2−•. However, other members of the NOX family generate much less O2−• and hence H2O2, and their activity is difficult to distinguish from other sources of these reactive species. In addition, O2−• and H2O2 are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O2−• and H2O2 in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives. [Display omitted] ► NOX enzymes have the sole known function to generate reactive oxygen species (ROS). ► Distinguishing NOX from other sources of ROS requires careful experimentation. ► This review gives a snapshot of methods used to detect NOX-derived ROS in tissues. ► Promising new technologies are discussed for the measurement of NOX activity.
AbstractList NADPH oxidases (NOX) are superoxide anion radical (O₂ ⁻•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O₂ ⁻•. NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O₂ ⁻• or its direct dismutation product hydrogen peroxide (H₂O₂) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O₂ ⁻•. However, other members of the NOX family generate much less O₂ ⁻• and hence H₂O₂, and their activity is difficult to distinguish from other sources of these reactive species. In addition, O₂ ⁻• and H₂O₂ are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O₂ ⁻• and H₂O₂ in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives.
NADPH oxidases (NOX) are superoxide anion radical (O(2)(-•))-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O(2)(-•). NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O(2)(-•) or its direct dismutation product hydrogen peroxide (H(2)O(2)) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O(2)(-•). However, other members of the NOX family generate much less O(2)(-•) and hence H(2)O(2), and their activity is difficult to distinguish from other sources of these reactive species. In addition, O(2)(-•) and H(2)O(2) are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O(2)(-•) and H(2)O(2) in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives.
NADPH oxidases (NOX) are superoxide anion radical (O2−•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O2−•. NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O2−• or its direct dismutation product hydrogen peroxide (H2O2) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O2−•. However, other members of the NOX family generate much less O2−• and hence H2O2, and their activity is difficult to distinguish from other sources of these reactive species. In addition, O2−• and H2O2 are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O2−• and H2O2 in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives. [Display omitted] ► NOX enzymes have the sole known function to generate reactive oxygen species (ROS). ► Distinguishing NOX from other sources of ROS requires careful experimentation. ► This review gives a snapshot of methods used to detect NOX-derived ROS in tissues. ► Promising new technologies are discussed for the measurement of NOX activity.
NADPH oxidases (NOX) are superoxide anion radical (O(2)(-•))-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O(2)(-•). NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O(2)(-•) or its direct dismutation product hydrogen peroxide (H(2)O(2)) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O(2)(-•). However, other members of the NOX family generate much less O(2)(-•) and hence H(2)O(2), and their activity is difficult to distinguish from other sources of these reactive species. In addition, O(2)(-•) and H(2)O(2) are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O(2)(-•) and H(2)O(2) in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives.NADPH oxidases (NOX) are superoxide anion radical (O(2)(-•))-generating enzymes. They form a family of seven members, each with a specific tissue distribution. They function as electron transport chains across membranes, using NADPH as electron donor to reduce molecular oxygen to O(2)(-•). NOX have multiple biological functions, ranging from host defense to inflammation and cellular signaling. Measuring NOX activity is crucial in understanding the roles of these enzymes in physiology and pathology. Many of the methods used to measure NOX activity are based on the detection of small molecules that react with NOX-generated O(2)(-•) or its direct dismutation product hydrogen peroxide (H(2)O(2)) to form fluorescent, luminescent, or colored products. Initial techniques were developed to measure the activity of the phagocyte isoform NOX2 during the oxidative burst of stimulated polymorphonuclear leukocytes, which generate large quantities of O(2)(-•). However, other members of the NOX family generate much less O(2)(-•) and hence H(2)O(2), and their activity is difficult to distinguish from other sources of these reactive species. In addition, O(2)(-•) and H(2)O(2) are reactive molecules and most probes are prone to artifacts and therefore should be used with appropriate controls and the data carefully interpreted. This review gives an overview of current methods used to measure NOX activity and NOX-derived O(2)(-•) and H(2)O(2) in cells, tissues, isolated systems, and living organisms, describing the advantages and caveats of many established methods with emphasis on more recent technologies and future perspectives.
Author Stocker, Roland
Maghzal, Ghassan J.
Jaquet, Vincent
Krause, Karl-Heinz
Author_xml – sequence: 1
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  surname: Maghzal
  fullname: Maghzal, Ghassan J.
  organization: Centre for Vascular Research, School of Medical Sciences (Pathology) and Bosch Institute, The University of Sydney, Medical Foundation Building, K25, NSW 2006, Australia
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  givenname: Karl-Heinz
  surname: Krause
  fullname: Krause, Karl-Heinz
  organization: Department of Pathology and Immunology, Geneva Medical Faculty and Department of Genetic and Laboratory Medicine, Geneva University Hospitals Centre Médical Universitaire 1, rue Michel-Servet, 1211 Geneva 4, Switzerland
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  fullname: Stocker, Roland
  organization: Centre for Vascular Research, School of Medical Sciences (Pathology) and Bosch Institute, The University of Sydney, Medical Foundation Building, K25, NSW 2006, Australia
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  surname: Jaquet
  fullname: Jaquet, Vincent
  email: Vincent.Jaquet@unige.ch
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/22982596$$D View this record in MEDLINE/PubMed
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ISSN 0891-5849
1873-4596
IngestDate Sun Sep 28 02:06:47 EDT 2025
Sun Sep 28 05:58:33 EDT 2025
Thu Apr 03 06:59:50 EDT 2025
Tue Nov 18 22:15:33 EST 2025
Sat Nov 29 07:10:43 EST 2025
Fri Feb 23 02:25:15 EST 2024
IsDoiOpenAccess false
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Issue 10
Keywords Oxidative stress
Reactive oxygen species
O2
Hydrogen peroxide
E
MRI
Fluorescence
H2O2
Superoxide anion radical
TPO
L-012
NADPH oxidase
DUOX
ONOO
PG1
GSH
Probes
DCFH
HPLC
CGD
NBT
ESR
MS
SOD
WST-1
ER
AGT
MCLA
cpYFP
HOCl
NOX
ROS
DPI
HE
2-OH-E
Mass spectrometry
PET
Language English
License Copyright © 2012 Elsevier Inc. All rights reserved.
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PublicationTitle Free radical biology & medicine
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Snippet NADPH oxidases (NOX) are superoxide anion radical (O2−•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution....
NADPH oxidases (NOX) are superoxide anion radical (O(2)(-•))-generating enzymes. They form a family of seven members, each with a specific tissue distribution....
NADPH oxidases (NOX) are superoxide anion radical (O₂ ⁻•)-generating enzymes. They form a family of seven members, each with a specific tissue distribution....
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elsevier
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StartPage 1903
SubjectTerms Biological Assay
Biosensing Techniques
Cell-Free System
Cytochromes c - chemistry
Electron Spin Resonance Spectroscopy
electron transfer
enzymes
Fluorescence
HPLC
Hydrogen peroxide
inflammation
Mass spectrometry
NADH, NADPH Oxidoreductases - metabolism
NADP (coenzyme)
NADPH oxidase
neutrophils
NOX
Oxidation-Reduction
Oxidative stress
oxygen
Probes
Reactive oxygen species
Reactive Oxygen Species - analysis
superoxide anion
Superoxide anion radical
Superoxides - analysis
Tetrazolium Salts - chemistry
tissue distribution
Title Detection of reactive oxygen species derived from the family of NOX NADPH oxidases
URI https://dx.doi.org/10.1016/j.freeradbiomed.2012.09.002
https://www.ncbi.nlm.nih.gov/pubmed/22982596
https://www.proquest.com/docview/1151036389
https://www.proquest.com/docview/1678521021
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