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The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels

Abstract

The enzyme NADPH oxidase in phagocytes is important in the body's defence against microbes: it produces superoxide anions (O2-, precursors to bactericidal reactive oxygen species1). Electrons move from intracellular NADPH, across a chain comprising FAD (flavin adenine dinucleotide) and two haems, to reduce extracellular O2 to O2-. NADPH oxidase is electrogenic2, generating electron current (Ie) that is measurable under voltage-clamp conditions3,4. Here we report the complete current–voltage relationship of NADPH oxidase, the first such measurement of a plasma membrane electron transporter. We find that Ie is voltage-independent from -100 mV to >0 mV, but is steeply inhibited by further depolarization, and is abolished at about +190 mV. It was proposed that H+ efflux2 mediated by voltage-gated proton channels5,6 compensates Ie, because Zn2+ and Cd2+ inhibit both H+ currents7,8,9 and O2- production10. Here we show that COS-7 cells transfected with four NADPH oxidase components11, but lacking H+ channels12, produce O2- in the presence of Zn2+ concentrations that inhibit O2- production in neutrophils and eosinophils. Zn2+ does not inhibit NADPH oxidase directly, but through effects on H+ channels. H+ channels optimize NADPH oxidase function by preventing membrane depolarization to inhibitory voltages.

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Figure 1: Current–voltage relationship for NADPH-oxidase-mediated electron current (Ie) in human eosinophils.
Figure 2: Factors affecting superoxide production.

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Acknowledgements

We thank A. R. Cross and L. L. Thomas for discussions, and T. Iastrebova and J. Murphy for technical assistance. This work was supported in part by the Heart, Lung and Blood Institute of the National Institutes of Health (T.E.D.).

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Correspondence to Thomas E. DeCoursey.

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DeCoursey, T., Morgan, D. & Cherny, V. The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels. Nature 422, 531–534 (2003). https://doi.org/10.1038/nature01523

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