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Methods for Detection of NOX-Derived Superoxide Radical Anion and Hydrogen Peroxide in Cells

Fiona Augsburger1, Aleksandra Filippova1, Vincent Jaquet2,3

  • 1Faculty of Medicine, Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)
|June 8, 2019
PubMed

Insights

Researchers developed two cellular assays to detect extracellular superoxide (O2ċ-) and hydrogen peroxide (H2O2) produced by NADPH oxidases (NOX). These assays aid in studying NOX functions and developing new treatments for diseases linked to ROS.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Enzymology

Background:

  • NADPH oxidases (NOX) are key enzymes producing reactive oxygen species (ROS).
  • The NOX family includes NOX1-5 and DUOX1-2, generating superoxide (O2ċ-) or hydrogen peroxide (H2O2).
  • NOX-derived ROS play critical roles in physiological processes and diseases like diabetes and ischemia.

Purpose of the Study:

  • To develop and describe two novel cellular assays.
  • To detect extracellular superoxide radical anion (O2ċ-) and hydrogen peroxide (H2O2).
  • To facilitate research on cells overexpressing specific NOX isoforms and their subunits.

Main Methods:

  • Development of two distinct cellular assays.
  • Utilizing cells engineered to overexpress specific NOX isoforms and subunits.
  • Quantification of extracellular O2ċ- and H2O2 production.

Main Results:

  • Successful establishment of assays for detecting extracellular O2ċ- and H2O2.
  • Demonstrated ability to measure ROS production from specific NOX isoforms.
  • Provided a tool for investigating NOX enzyme activity in cellular contexts.

Conclusions:

  • The developed assays are valuable tools for studying NOX enzymology.
  • These assays can advance research into the roles of NOX-derived ROS in health and disease.
  • Facilitates the screening of NOX inhibitors for therapeutic development.
Keywords:
Amplex redCellular assayHydrogen peroxideNADPH oxidasePeroxidaseSuperoxide radical anionWST-1

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