The gp91phox component of NADPH oxidase is not the voltage-gated proton channel in phagocytes, but it helps.
During the "respiratory burst," the NADPH oxidase complex of phagocytes produces reactive oxygen species that kill bacteria and other invaders (Babior, B. M. (1999) Blood 93, 1464-1476). Electron efflux through NADPH oxidase is electrogenic (Henderson, L. M., Chappell, J. B., and Jones, O. T. G. (1987) Biochem. J. 246, 325-329) and is compensated by H(+) efflux through proton channels that reportedly are contained within the gp91(phox) subunit of NADPH oxidase. To test whether gp91(phox) functions as a proton channel, we studied H(+) currents in granulocytes from X-linked chronic granulomatous disease patients lacking gp91(phox) (X-CGD), the human myelocytic PLB-985 cell line, PLB-985 cells in which gp91(phox) was knocked out by gene targeting (PLB(KO)), and PLB-985 knockout cells re-transfected with gp91(phox) (PLB(91)). H(+) currents in unstimulated PLB(KO) cells had amplitude and gating kinetics similar to PLB(91) cells. Furthermore, stimulation with the phorbol ester phorbol 12-myristate 13-acetate increased H(+) currents to a similar extent in X-CGD, PLB(KO), and PLB(91) cells. Thus, gp91(phox) is not the proton channel in unstimulated phagocytes and does not directly mediate the increase of proton conductance during the respiratory burst. Changes in H(+) channel gating kinetics during NADPH oxidase activity are likely crucial to the activation of H(+) flux during the respiratory burst.
DeCoursey
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Department of Molecular Biophysics and Physiology, Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612, USA. tdecours@rush.edu
Cherny
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Morgan
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Katz
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Dinauer
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D002273:Carcinogens; D002460:Cell Line; D004594:Electrophysiology; D006105:Granulomatous Disease, Chronic / Q000235:genetics; D006801:Humans; D006859:Hydrogen / Q000378:metabolism; D007477:Ions; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000502:physiology*; D009154:Mutation; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000737:chemistry*; D010586:Phagocytes / Q000378:metabolism*; D011522:Protons; D016897:Respiratory Burst; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism; D019076:Transgenes
D002273:Carcinogens; D007477:Ions; D008562:Membrane Glycoproteins; D011522:Protons; D006859:Hydrogen; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate