Cited by 632Redox biology and oxidative stressNitric Oxide and Endothelin EffectsNeutrophil, Myeloperoxidase and Oxidative Mechanisms

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

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Serrander, Lena, Cartier, Laetitia, Bedard, Karen, Banfi, Botond, Lardy, Bernard, Plastre, Olivier, Sienkiewicz, Andrzej, Fórró, Lászlo, Schlegel, Werner, Krause, Karl-Heinz·2007·Biochemical Journal

DOI 10.1042/bj20061903PMC PMC1948990

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Abstract

NOX4 is an enigmatic member of the NOX (NADPH oxidase) family of ROS (reactive oxygen species)-generating NADPH oxidases. NOX4 has a wide tissue distribution, but the physiological function and activation mechanisms are largely unknown, and its pharmacology is poorly understood. We have generated cell lines expressing NOX4 upon tetracycline induction. Tetracycline induced a rapid increase in NOX4 mRNA (1 h) followed closely (2 h) by a release of ROS. Upon tetracycline withdrawal, NOX4 mRNA levels and ROS release decreased rapidly (<24 h). In membrane preparations, NOX4 activity was selective for NADPH over NADH and did not require the addition of cytosol. The pharmacological profile of NOX4 was distinct from other NOX isoforms: DPI (diphenyleneiodonium chloride) and thioridazine inhibited the enzyme efficiently, whereas apocynin and gliotoxin did not (IC(50)>100 muM). The pattern of NOX4-dependent ROS generation was unique: (i) ROS release upon NOX4 induction was spontaneous without need for a stimulus, and (ii) the type of ROS released from NOX4-expressing cells was H(2)O(2), whereas superoxide (O(2)(-)) was almost undetectable. Probes that allow detection of intracellular O(2)(-) generation yielded differential results: DHE (dihydroethidium) fluorescence and ACP (1-acetoxy-3-carboxy-2,2,5,5-tetramethylpyrrolidine) ESR measurements did not detect any NOX4 signal, whereas a robust signal was observed with NBT. Thus NOX4 probably generates O(2)(-) within an intracellular compartment that is accessible to NBT (Nitro Blue Tetrazolium), but not to DHE or ACP. In conclusion, NOX4 has a distinct pharmacology and pattern of ROS generation. The close correlation between NOX4 mRNA and ROS generation might hint towards a function as an inducible NOX isoform.

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Authors: Serrander, L., Cartier, L., Bedard, K., Banfi, B., Lardy, B., Plastre, O., Sienkiewicz, A., Fórró, L., Schlegel, W., & Krause, K.-H.
Affiliations: University of Geneva (CH)
Department of Anatomy and Cell Biology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, U.S.A.
Hôpital Albert Michallon (FR)
École Polytechnique Fédérale de Lausanne (CH)
Institute of Physics of Complex Matter, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland
Corresponding author: Serrander, Lena
Schlegel, Werner
ORCID: Serrander, Lena
Bedard, Karen
Sienkiewicz, Andrzej
Krause, Karl-Heinz
Venue: Biochemical Journal
Publisher: Portland Press Ltd.
Published: 2007-07-26
Volume: 406
Issue: 1
Pages: 105-114
ISSN: 0264-6021 (ISSN-L)
1470-8728
DOI: 10.1042/bj20061903
PMC: PMC1948990
OpenAlex: W1998073026
Cited by: 627
References: 49
Citations by year: 2026: 9 · 2025: 19 · 2024: 19 · 2023: 25 · 2022: 22 · 2021: 37 · 2020: 30 · 2019: 36 · 2018: 31 · 2017: 29 · 2016: 33 · 2015: 47 · 2014: 51 · 2013: 50 · 2012: 43
FWCI: 13.23
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: Biochemical Journal (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
HAL (submittedVersion)
HAL (submittedVersion, other-oa)
Trinity's Access to Research Output (TARA) (Trinity College Dublin) (submittedVersion, other-oa)
Archive ouverte UNIGE (University of Geneva) (submittedVersion)
Archive ouverte UNIGE (University of Geneva) (submittedVersion, cc-by)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
Keywords: NOX4, NADPH oxidase, reactive oxygen species, hydrogen peroxide, superoxide, mRNA expression, tetracycline-inducible expression, diphenyleneiodonium, thioridazine, apocynin, gliotoxin, nitroblue tetrazolium reduction
MeSH terms: NADPH Oxidase 4, Animals, Cell Line, Cell Membrane, Cytosol, Electron Spin Resonance Spectroscopy, Enzyme Induction, Enzyme Inhibitors, Ethidium, Humans, Hydrogen Peroxide, Mitochondria, NAD, NADP, Nitroblue Tetrazolium, RNA, Messenger, Superoxides, Tetracycline, Time Factors, Gene Expression Regulation, Enzymologic, Reactive Oxygen Species, NADPH Oxidases, Mice

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

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