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NOX enzymes and the biology of reactive oxygen

Key Points

  • The phagocyte NADPH oxidase (Phox) is a well-characterized enzyme that generates high levels of superoxide and secondary oxidants in phagocytes as part of the armoury of microbicidal mechanisms by which these cells function in host defence.

  • The catalytic moiety of Phox is the membrane-associated flavocytochrome gp91phox, which is inactive in resting phagocytes, but becomes activated in the phagosomal membrane by assembly with regulatory subunits such as p47phox, p67phox and RAC.

  • A new family of homologues of gp91phox, the NADPH oxidase (NOX)/dual oxidase (DUOX) family, which now contains seven members, has recently been described, and these are expressed in various cell types, including the epithelium, smooth-muscle cells and the endothelium. These enzymes 'deliberately' generate superoxide and secondarily produce other reactive oxygen species (ROS), including hydrogen peroxide (H2O2).

  • The NOX enzymes have been proposed to function to generate ROS as mediators of signal transduction related to growth, angiogenesis and apoptosis. In addition, circumstantial evidence indicates that NOX enzymes might in some cases function in innate immunity in barrier cells, such as the colon epithelium, in a manner that is analogous to Phox.

  • The DUOX enzymes are dual function enzymes, containing not only an ROS-generating domain that is homologous to gp91phox, but also a peroxidase domain that can use the H2O2 produced by the gp91phox-homology domain to carry out oxidations of other substrates. A DUOX enzyme in the thyroid has been shown to participate in thyroid-hormone biosynthesis.

  • A Duox enzyme in Caenorhabditis elegans catalyses the crosslinking of tyrosine residues in the cuticle — an exoskeletal structure in nematode worms. The structure and function of DUOX enzymes implies a general function in the oxidative modification of the extracellular matrix or other extracellular molecules.

  • The NOX/DUOX family is implicated in various pathological conditions, including atherosclerosis, hypertension, cancer and endocrine disorders. These enzymes provide an attractive target for new therapeutic agents.

  • Our understanding of the enzymology and subunit composition of the NOX/DUOX family of enzymes is evolving rapidly. However, definitive information regarding the biological roles of these enzymes is largely lacking and will require the development of animal model systems (Drosophila and knockout mice, for example).

Abstract

Professional phagocytes generate high levels of reactive oxygen species (ROS) using a superoxide-generating NADPH oxidase as part of their armoury of microbicidal mechanisms. The multicomponent phagocyte oxidase (Phox), which has been well characterized over the past three decades, includes the catalytic subunit gp91phox. Lower levels of ROS are seen in non-phagocytic cells, but are usually thought to be 'accidental' byproducts of aerobic metabolism. The discovery of a family of superoxide-generating homologues of gp91phox has led to the concept that ROS are 'intentionally' generated in these cells with distinctive cellular functions related to innate immunity, signal transduction and modification of the extracellular matrix.

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Figure 1: Reactive oxygen species.
Figure 2: Transmembrane topology and domain structure of NOX and DUOX enzymes.
Figure 3: Domain structure of regulatory proteins for NOX enzymes.
Figure 4: Activation of reactive oxygen species (ROS) generation by assembly of Phox regulatory proteins in phagocytes.

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Acknowledgements

I am grateful to D. Ritsick for suggesting important concepts regarding the biology and signalling of reactive oxygen species that have been incorporated into this article.

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DATABASES

LocusLink

DUOX1

DUOX2

gp91phox

KGF-α

NOX1

NOX3

NOX4

NOX5

NOXA1

NOXO1

p22phox

p40phox

p47phox

p67phox

PTEN

RAC1

RAC2

RhoGDI

Sod2

TLR4

FURTHER INFORMATION

Website for Second International Conference on NADPH Oxidases

David Lambeth's Lab

Glossary

SUPEROXIDE

The one-electron-reduced form of molecular oxygen.

PEROXIDASES

Haeme-containing enzymes that metabolize hydrogen peroxide with the concomitant oxidation of a co-substrate, such as chloride in the case of myeloperoxidase.

NADPH

The reduced form of nicotinamide adenine dinucleotide phosphate. This co-enzyme serves as an electron donor for various biochemical reactions.

RESPIRATORY BURST

The large increase in oxygen consumption and reactive oxygen generation that accompanies exposure of neutrophils to microorganisms and/or inflammatory mediators.

FLAVOPROTEIN

A protein or enzyme that contains a flavin co-enzyme such as flavin adenine dinucleotide (FAD).

GASTRIC PIT CELLS

Specialized secretory epithelial cells found in the stomach.

SENESCENCE

A cellular phenotype typically seen in primary cells that have undergone many cell divisions, and typified by altered morphology and an inability to continue to divide. Cell senescence is also seen in cells that have been exposed to oxidants and other stresses.

NUDE MICE

A line of mice in which lymphocyte immune function is compromised due to the congenital absence of the thymus. These mice are frequently used for tumour studies, because there is no rejection of the tumour.

THYROGLOBULIN

The protein that serves as a precursor to thyroid hormone. Tyrosine residues in thyroglobulin are first iodinated in a reaction catalysed by thyroid peroxidase, and are then crosslinked to form the precursor of thyroid hormone. Proteolysis of thyroglobulin then releases the thyroid hormone from the protein structure.

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Lambeth, J. NOX enzymes and the biology of reactive oxygen. Nat Rev Immunol 4, 181–189 (2004). https://doi.org/10.1038/nri1312

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