Cited by 145Redox biology and oxidative stressNitric Oxide and Endothelin EffectsNeutrophil, Myeloperoxidase and Oxidative Mechanisms

Identification of Structural Elements in Nox1 and Nox4 Controlling Localization and Activity

Helmcke, Ina, Heumüller, Sabine, Tikkanen, Ritva, Schröder, Katrin, Brandes, Ralf P.·2009·Antioxidants & Redox Signaling

Abstract

Nox NADPH oxidases differ in their mode of activation, subcellular localization, and physiological function. Nox1 releases superoxide anions (O(2)(-)) and depends on cytosolic activator proteins, whereas Nox4 extracellularly releases hydrogen peroxide (H(2)O(2)), and its activity does not require cotransfection of additional proteins. We constructed chimeric proteins consisting of Nox1 and Nox4 expressed in HEK293 cells. When the cytosolic tail of Nox4 was fused with the transmembrane part of Nox1, Nox1 became constitutively active. The reciprocal construct was inactive, suggesting that cytosolic subunit-dependent activation requires elements in the transmembrane loops. By TIRF-microscopy, Nox1 was observed in the plasma membrane, whereas Nox4 colocalized with proteins of the endoplasmic reticulum. Fusion proteins of Myc and Nox revealed that the N-terminal part of Nox1 but not Nox4 is cleaved. When the potential signal peptide of Nox4 was inserted into Nox1, plasma-membrane localization was lost, and the protein was retained in vesicle-like structures below the plasma membrane. The potential signal peptide of Nox1 failed to translocate Nox4 to the plasma membrane but switched the extracellularly detectable ROS from H(2)O(2) to O(2)(-). Thus, the very N-terminal part of Nox proteins determines subcellular localization and the ROS type released, whereas the cytosolic tail regulates activity.

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Crossref · 10.1089/ars.2008.2383

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Authors
Helmcke, I., Heumüller, S., Tikkanen, R., Schröder, K., & Brandes, R. P.
Affiliations
  • Goethe University Frankfurt (DE)
  • Institut für Kardiovaskuläre Physiologie, Goethe-Universität, Frankfurt am Main, Germany.
Corresponding author
Brandes, Ralf P.
Venue
Antioxidants & Redox Signaling
Published
2009-06
Volume
11
Issue
6
Pages
1279-1287
ISSN
  • 1523-0864 (ISSN-L)
  • 1557-7716
OpenAlex
W2058114038
Cited by
145
References
38
Citations by year
2026: 3 · 2024: 5 · 2023: 8 · 2022: 5 · 2021: 9 · 2020: 3 · 2019: 7 · 2018: 5 · 2017: 7 · 2016: 8 · 2015: 8 · 2014: 8 · 2013: 15 · 2012: 18
FWCI
4.93
Type
journal article
Language
en
Open access
closed
Locations
  • Antioxidants and Redox Signaling (publishedVersion)
  • PubMed (publishedVersion)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
  • Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
Keywords
NOX1, NOX4, NADPH oxidase, fusion protein, subcellular localization, superoxide, hydrogen peroxide, cytoplasmic tail, signal peptide, N-terminal cleavage, plasma membrane, endoplasmic reticulum, constitutive activation, reactive oxygen species
MeSH terms
NADPH Oxidase 1, NADPH Oxidase 4, Cell Line, Humans, Microscopy, Fluorescence, Models, Biological, Recombinant Proteins, Blotting, Western, Reactive Oxygen Species, Microscopy, Confocal, NADPH Oxidases, Protein Sorting Signals

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