NADPH Oxidases

1. Front Matter

   Pages i-xv

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2. The NADPH Oxidase Family: Overviews

   1. Front Matter

      Pages 1-1

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   2. Intersecting Stories of the Phagocyte NADPH Oxidase and Chronic Granulomatous Disease

      • William M. Nauseef, Robert A. Clark

      Pages 3-16

   3. Mammalian NADPH Oxidases

      • Hélène Buvelot, Vincent Jaquet, Karl-Heinz Krause

      Pages 17-36

3. Biochemistry and Model Systems

   1. Front Matter

      Pages 37-37

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   2. Enhanced Immunoaffinity Purification of Human Neutrophil Flavocytochrome B for Structure Determination by Electron Microscopy

      • Algirdas J. Jesaitis, Marcia Riesselman, Ross M. Taylor, Susan Brumfield

      Pages 39-59

   3. Purification and Characterization of DUOX Peroxidase Homology Domains (PHDs)

      • Jennifer L. Meitzler

      Pages 61-74

   4. A Close-Up View of the Impact of Arachidonic Acid on the Phagocyte NADPH Oxidase

      • Tania Bizouarn, Hager Souabni, Xavier Serfaty, Aicha Bouraoui, Rawand Masoud, Gilda Karimi et al.

      Pages 75-101

   5. NOX5 Cell-Free Assay for the High-Throughput Screening of Small Molecules

      • Fiona Augsburger, Delphine Rasti, Yves Cambet, Vincent Jaquet

      Pages 103-111

   6. Spectroscopy of NOX Protein Family Members

      • Yoko Nakano, William M. Nauseef

      Pages 113-120

   7. Soluble Regulatory Proteins for Activation of NOX Family NADPH Oxidases

      • Hideki Sumimoto, Reiko Minakami, Kei Miyano

      Pages 121-137

   8. Insights into the NOX NADPH Oxidases Using Heterologous Whole Cell Assays

      • Mary C. Dinauer

      Pages 139-151

   9. The X-CGD PLB-985 Cell Model for NOX2 Structure-Function Analysis

      • Sylvain Beaumel, Marie José Stasia

      Pages 153-171

   10. Functional Characterization of DUOX Enzymes in Reconstituted Cell Models

       • Agnieszka Korzeniowska, Ágnes P. Donkó, Stanislas Morand, Thomas L. Leto

       Pages 173-190

   11. Guidelines for the Detection of NADPH Oxidases by Immunoblot and RT-qPCR

       • Becky A. Diebold, S. Garrett Wilder, Xavier De Deken, Jennifer L. Meitzler, James H. Doroshow, James W. McCoy et al.

       Pages 191-229

4. Detection of ROS

   1. Front Matter

      Pages 231-231

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

      • Fiona Augsburger, Aleksandra Filippova, Vincent Jaquet

      Pages 233-241

   3. HPLC-Based Monitoring of Oxidation of Hydroethidine for the Detection of NADPH Oxidase-Derived Superoxide Radical Anion

      • Jacek Zielonka, Monika Zielonka, Balaraman Kalyanaraman

      Pages 243-258

   4. Visualization of Intracellular Hydrogen Peroxide with the Genetically Encoded Fluorescent Probe HyPer in NIH-3T3 Cells

      • Yulia G. Ermakova, Nataliya M. Mishina, Carsten Schultz, Vsevolod V. Belousov

      Pages 259-274

   5. Imaging Intracellular H2O2 with the Genetically Encoded PerFRET and OxyFRET Probes

      • Balázs Enyedi, Miklós Geiszt

      Pages 275-282

This detailed volume explores the NADPH oxidase family of enzymes in human physiology and genetic disease, in which early discoveries represent prime examples of the finest translational “from bed to bench and back” studies. Methods are included for testing assembly and function of multicomponent oxidase complexes and for analyzing reactive oxygen species (ROS) generation in different systems by various means, while addressing pitfalls of ROS probes currently being used, as well as protocols on NADPH oxidase regulation and their function in cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and cutting-edge, NADPH Oxidases: Methods and Protocols will aid researchers working with the NOX/DUOX family in continuing and expanding upon their vital research.

• NOX/DUOX family
• Multicomponent oxidase complexes
• Genetic diseases
• Reactive oxygen species
• Oxidase regulation
• Disease modeling

• Conway Institute, School of Medicine, University College Dublin, Dublin, Ireland

  Ulla G. Knaus

• Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, USA

  Thomas L. Leto

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