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Updated: May 10, 2026
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Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
Peroxiredoxin-6 and NADPH oxidase activity
1Department of Pediatrics, University of Colorado Denver, Anschutz Medical Campus, Aurora, Colorado, USA. daniel.ambruso@ucdenver.edu
Methods in Enzymology
|July 9, 2013
Summary
Peroxiredoxin 6 (Prdx6) aids neutrophil function by supporting NADPH oxidase (Nox2) activity. Researchers developed methods to study Prdx6
Area of Science:
- Biochemistry
- Cell Biology
- Immunology
Background:
- Peroxiredoxins (Prdxs) are cysteine-based antioxidant enzymes crucial for cellular redox homeostasis.
- Prdx6, a 1-cys Prdx, is implicated in neutrophil function and associates with NADPH oxidase (Nox2).
Purpose of the Study:
- To detail methods for assessing Prdx6 activity and its influence on Nox2.
- To describe techniques for modulating Prdx6 expression in myeloid cells to study its role in Nox2 activity.
Main Methods:
- Enzyme activity assays for determining Prdx6's catalytic function.
- Cell-free systems utilizing SDS-activated NADPH oxidase to evaluate Prdx6's effect on Nox2.
- RNA interference (siRNA and shRNA) for suppressing Prdx6 expression in K562 and cultured myeloid cells.
Main Results:
- Established protocols for quantifying Prdx6 activity.
- Demonstrated an approach to assess Prdx6's impact on Nox2 in a cell-free system.
- Outlined methods for gene silencing of Prdx6 in relevant cell types.
Conclusions:
- The described methods enable comprehensive investigation of Prdx6's role in neutrophil function and Nox2 regulation.
- These techniques facilitate exploration of the biochemical mechanisms underlying Prdx6's interaction with the NADPH oxidase system.
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