{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,14]],"date-time":"2026-03-14T03:55:44Z","timestamp":1773460544107,"version":"3.50.1"},"reference-count":43,"publisher":"Rockefeller University Press","issue":"6","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1999,12,1]]},"abstract":"<jats:p>Expression of gp91-phox in Chinese hamster ovary (CHO91) cells is correlated with the presence of a voltage-gated H+ conductance. As one component of NADPH oxidase in neutrophils, gp91-phox is responsible for catalyzing the production of superoxide (O2\u00b72). Suspensions of CHO91 cells exhibit arachidonate-activatable H+ fluxes (Henderson, L.M., G. Banting, and J.B. Chappell. 1995. J. Biol. Chem. 270:5909\u20135916) and we now characterize the electrical properties of the pathway. Voltage-gated currents were recorded from CHO91 cells using the whole-cell configuration of the patch-clamp technique under conditions designed to exclude a contribution from ions other than H+. As in other voltage-gated proton currents (Byerly, L., R. Meech, and W. Moody. 1984. J. Physiol. 351:199\u2013216; DeCoursey, T.E., and V.V. Cherny. 1993. Biophys. J. 65:1590\u20131598), a lowered external pH (pHo) shifted activation to more positive voltages and caused the tail current reversal potential to shift in the manner predicted by the Nernst equation. The outward currents were also reversibly inhibited by 200 \u03bcM zinc. Voltage-gated currents were not present immediately upon perforating the cell membrane, but showed a progressive increase over the first 10\u201320 min of the recording period. This time course was consistent with a gradual shift in activation to more negative potentials as the pipette solution, pH 6.5, equilibrated with the cell contents (reported by Lucifer yellow included in the patch pipette). Use of the pH-sensitive dye 2\u20327\u2032 bis-(2-carboxyethyl)-5(and 6) carboxyfluorescein (BCECF) suggested that the final intracellular pH (pHi) was \u223c6.9, as though pHi was largely determined by endogenous cellular regulation. Arachidonate (20 \u03bcM) increased the amplitude of the currents by shifting activation to more negative voltages and by increasing the maximally available conductance. Changes in external Cl\u2212 concentration had no effect on either the time scale or the appearance of the currents. Examination of whole cell currents from cells expressing mutated versions of gp91-phox suggest that: (a) voltage as well as arachidonate sensitivity was retained by cells with only the NH2-terminal 230 amino acids, (b) histidine residues at positions 111, 115, and 119 on a putative membrane-spanning helical region of the protein contribute to H+ permeation, (c) histidine residues at positions 111 and 119 may contribute to voltage gating, (d) the histidine residue at position 115 is functionally important for H+ selectivity. Mechanisms of H+ permeation through gp91-phox include the possible protonation\/deprotonation of His-115 as it is exposed alternatively to the interior and exterior faces of the cell membrane (see Starace, D.M., E. Stefani, and F. Bezanilla. 1997. Neuron. 19:1319\u20131327) and the transfer of protons across an \u201cH-X-X-X-H-X-X-X-H\u201d motif lining a conducting pore.<\/jats:p>","DOI":"10.1085\/jgp.114.6.771","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T16:50:20Z","timestamp":1027702220000},"page":"771-786","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":56,"title":["Evidence That the Product of the Human X-Linked Cgd Gene, Gp91-<i>phox<\/i>, Is a Voltage-Gated H+ Pathway"],"prefix":"10.1085","volume":"114","author":[{"given":"Lydia M.","family":"Henderson","sequence":"first","affiliation":[{"name":"aFrom the Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol, United Kingdom BS8 1TD"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert W.","family":"Meech","sequence":"additional","affiliation":[{"name":"bFrom the Department of Physiology, School of Medical Sciences, University of Bristol, Bristol, United Kingdom BS8 1TD"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[1999,11,15]]},"reference":[{"key":"2023072918145169100_ArmstrongandBezanilla1974","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1085\/jgp.63.5.533","article-title":"Charge movement associated with the opening and closing of the activation gates of the Na channels","volume":"63","author":"Armstrong","year":"1974","journal-title":"J. Gen. Physiol."},{"key":"2023072918145169100_BarishandBaud1984","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1113\/jphysiol.1984.sp015289","article-title":"A voltage gated hydrogen ion current in the oocytes membrane of the axolotol, Ambystoma","volume":"352","author":"Barish","year":"1984","journal-title":"J. Physiol"},{"key":"2023072918145169100_Bernheimetal1993","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1113\/jphysiol.1993.sp019860","article-title":"A voltage-dependent proton current in cultured human skeletal muscle myotubes","volume":"470","author":"Bernheim","year":"1993","journal-title":"J. Physiol"},{"key":"2023072918145169100_Byerlyetal1984","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1113\/jphysiol.1984.sp015241","article-title":"Rapidly activating hydrogen ion currents in perfused neurones of the snail, Lymnaea stagnalis","volume":"351","author":"Byerly","year":"1984","journal-title":"J. Physiol."},{"key":"2023072918145169100_ByerlyandMoody1986","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1113\/jphysiol.1986.sp016165","article-title":"Membrane currents of internally perfused neurons of the snail, Lymnaea stagnalis, at low intracellular pH","volume":"376","author":"Byerly","year":"1986","journal-title":"J. Physiol"},{"key":"2023072918145169100_ByerlyandSuen1989","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1113\/jphysiol.1989.sp017642","article-title":"Characterization of proton currents in neurones of the snail, Lymnaea stagnalis","volume":"413","author":"Byerly","year":"1989","journal-title":"J. Physiol."},{"key":"2023072918145169100_Chernyetal1996","first-page":"A77","article-title":"Proton and chloride currents in Chinese hamster ovary cells","volume":"70","author":"Cherny","year":"1996","journal-title":"Biophys. J."},{"key":"2023072918145169100_DeCourseyandCherny1998","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1085\/jgp.112.4.503","article-title":"Temperature dependence of voltage-gated H+ currents in human neutrophils, rat aveolar epithelial cells, and mammalian phagocytes","volume":"112","author":"DeCoursey","year":"1998","journal-title":"J. Gen. Physiol"},{"key":"2023072918145169100_DeCourseyandCherny1996","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/S0006-3495(96)79215-9","article-title":"Effects of buffer concentration on voltage-gated H+ currentsdoes diffusion limit the conductance?","volume":"71","author":"DeCoursey","year":"1996","journal-title":"Biophys. J"},{"key":"2023072918145169100_DeCourseyandCherny1993","doi-asserted-by":"crossref","first-page":"1590","DOI":"10.1016\/S0006-3495(93)81198-6","article-title":"Potential, pH and arachidonate gate hydrogen ion currents in human neutrophils","volume":"65","author":"DeCoursey","year":"1993","journal-title":"Biophys. J."},{"key":"2023072918145169100_DeCourseyandCherny1994","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1007\/BF00235130","article-title":"Voltage-activated hydrogen ion currents","volume":"141","author":"DeCoursey","year":"1994","journal-title":"J. Membr. Biol"},{"key":"2023072918145169100_Demaurexetal1993","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1113\/jphysiol.1993.sp019723","article-title":"Proton currents in human granulocytesregulation by membrane potential and intracellular pH","volume":"466","author":"Demaurex","year":"1993","journal-title":"J. Physiol"},{"key":"2023072918145169100_Ederetal1995","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1007\/BF00373889","article-title":"Properties of voltage-gated currents of microglia developed using macrophage colony-stimulating factor","volume":"430","author":"Eder","year":"1995","journal-title":"Pfl\u00fcgers Arch."},{"key":"2023072918145169100_Edwards1994","doi-asserted-by":"crossref","DOI":"10.1017\/CBO9780511608421","volume-title":"Biochemistry and Physiology of the Neutrophil","author":"Edwards","year":"1994"},{"key":"2023072918145169100_Goldman1943","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1085\/jgp.27.1.37","article-title":"Potential, impedance and rectification in membranes","volume":"27","author":"Goldman","year":"1943","journal-title":"J. Gen. Physiol."},{"key":"2023072918145169100_Henderson1998","doi-asserted-by":"crossref","first-page":"33216","DOI":"10.1074\/jbc.273.50.33216","article-title":"Role of histidine identified by mutagenesis in the NADPH oxidase-associated H+ channel","volume":"273","author":"Henderson","year":"1998","journal-title":"J. Biol. Chem"},{"key":"2023072918145169100_Hendersonetal1995","doi-asserted-by":"crossref","first-page":"5909","DOI":"10.1074\/jbc.270.11.5909","article-title":"The arachidonate-activable, NADPH oxidase-associated H+ channel. Evidence that gp91-phox functions as an essential part of the channel","volume":"270","author":"Henderson","year":"1995","journal-title":"J. Biol. Chem."},{"key":"2023072918145169100_HendersonandChappell1996","first-page":"87","article-title":"NADPH oxidase of neutrophils Biochim","volume":"1273","author":"Henderson","year":"1996","journal-title":"Biophys. Acta."},{"key":"2023072918145169100_HendersonandChappell1992","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1042\/bj2830171","article-title":"The NADPH-oxidase-associated H+ channel is opened by arachidonate","volume":"283","author":"Henderson","year":"1992","journal-title":"Biochem. J."},{"key":"2023072918145169100_Hendersonetal1987","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1042\/bj2460325","article-title":"The superoxide-generating NADPH oxidase of human neutrophils is electrogenic and associated with an H+ channel","volume":"246","author":"Henderson","year":"1987","journal-title":"Biochem. J."},{"key":"2023072918145169100_Hendersonetal1988a","first-page":"285","article-title":"Superoxide generation by the electrogenic NADPH oxidase of human neutrophils is limited by the movement of a compensating charge","volume":"255","author":"Henderson","year":"1988","journal-title":"Biochem. J."},{"key":"2023072918145169100_Hendersonetal1988b","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1042\/bj2510563","article-title":"Internal pH changes associated with the activity of NADPH oxidase of human neutrophils. Further evidence for the presence of an H+ conducting channel","volume":"251","author":"Henderson","year":"1988","journal-title":"Biochem. J."},{"key":"2023072918145169100_Hendersonetal1997","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1042\/bj3250701","article-title":"The arachidonate-activatable, NADPH oxidase-associated H+ channel is contained within the multi-membrane-spanning N-terminal region of gp91-phox","volume":"325","author":"Henderson","year":"1997","journal-title":"Biochem. J."},{"key":"2023072918145169100_HodgkinandKatz1949","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1113\/jphysiol.1949.sp004310","article-title":"The effect of sodium ions on the electrical activity of the giant axon of the squid","volume":"108","author":"Hodgkin","year":"1949","journal-title":"J. Physiol."},{"key":"2023072918145169100_Kapusetal1993","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1085\/jgp.102.4.729","article-title":"A pH-sensitive and voltage-dependent proton conductance in the plasma membrane of macrophages","volume":"102","author":"Kapus","year":"1993","journal-title":"J. Gen. Physiol."},{"key":"2023072918145169100_Kapusetal1994","doi-asserted-by":"crossref","first-page":"4736","DOI":"10.1016\/S0021-9258(17)37606-8","article-title":"Arachidonate acid stimulates the plasma membrane H+ conductance of macrophages","volume":"269","author":"Kapus","year":"1994","journal-title":"J. Biol. Chem."},{"key":"2023072918145169100_Langton1993","first-page":"244P","article-title":"A versatile superfusion system suitable for whole-cell and exercised patch clamp experiments","volume":"467","author":"Langton","year":"1993","journal-title":"J. Physiol"},{"key":"2023072918145169100_Lukasetal1993","doi-asserted-by":"crossref","first-page":"C3","DOI":"10.1152\/ajpcell.1993.265.1.C3","article-title":"Proton conductance of the plasma membraneproperties, regulation, and functional role","volume":"265","author":"Lukas","year":"1993","journal-title":"Am. J. Physiol."},{"key":"2023072918145169100_Mahaut-Smith1989","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1242\/jeb.145.1.455","article-title":"The effect of zinc on calcium and hydrogen ion currents in intact snail neurones","volume":"145","author":"Mahaut-Smith","year":"1989","journal-title":"J. Exp. Biol."},{"key":"2023072918145169100_Mathiasetal1990","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1016\/S0006-3495(90)82418-8","article-title":"Limitations of the whole cell patch clamp technique in the control of intracellular concentrations","volume":"58","author":"Mathias","year":"1990","journal-title":"Biophys. J."},{"key":"2023072918145169100_MeechandThomas1980","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1113\/jphysiol.1980.sp013070","article-title":"Effect of measured calcium chloride injection on the membrane potential and internal pH of snail neurons","volume":"298","author":"Meech","year":"1980","journal-title":"J. Physiol"},{"key":"2023072918145169100_Olivaetal1988","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1016\/S0006-3495(88)83017-0","article-title":"Calculations of time constants for intracellular diffusion in whole cell patch clamp configuration","volume":"54","author":"Oliva","year":"1988","journal-title":"Biophys. J."},{"key":"2023072918145169100_PuschandNeher1988","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1007\/BF00582316","article-title":"Rates of diffusional exchange between small cells and a measuring patch pipette","volume":"411","author":"Pusch","year":"1988","journal-title":"Pfl\u00fcgers Arch"},{"key":"2023072918145169100_ReavesandBanting1994","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1016\/0014-5793(94)00813-2","article-title":"Overexpression of TGN38\/41 leads to mislocalisation of 7- adaptin","volume":"351","author":"Reaves","year":"1994","journal-title":"FEBS Lett."},{"key":"2023072918145169100_Roos1994","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1111\/j.1600-065X.1994.tb00850.x","article-title":"The genetic basis of chronic granulomatous disease","volume":"138","author":"Roos","year":"1994","journal-title":"Immunol. Rev."},{"key":"2023072918145169100_Royer-Pokoraetal1986","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1038\/322032a0","article-title":"Cloning the gene for an inherited human disorder\u2014chronic granulomatous disease\u2014on the basis of its chromosomal location","volume":"322","author":"Royer-Pokora","year":"1986","journal-title":"Nature."},{"key":"2023072918145169100_Schrenzeletal1998","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1038\/33725","article-title":"Electron currents generated by the human phagocyte NADPH oxidase","volume":"392","author":"Schrenzel","year":"1998","journal-title":"Nature."},{"key":"2023072918145169100_Schrenzeletal1996","doi-asserted-by":"crossref","first-page":"C1861","DOI":"10.1152\/ajpcell.1996.271.6.C1861","article-title":"Proton currents in human eosinophils","volume":"271","author":"Schrenzel","year":"1996","journal-title":"Am. J. Physiol"},{"key":"2023072918145169100_ShottonandWhite1989","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/0968-0004(89)90096-0","article-title":"Confocal scanning microscope3D biological imaging","volume":"14","author":"Shotton","year":"1989","journal-title":"TIBS (Trends Biochem. Sci.)."},{"key":"2023072918145169100_Staraceetal1997","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1016\/S0896-6273(00)80422-5","article-title":"Voltage-dependent proton transport by the voltage sensor of the Shaker K+ channel","volume":"19","author":"Starace","year":"1997","journal-title":"Neuron."},{"key":"2023072918145169100_Thomas1976","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1113\/jphysiol.1976.sp011305","article-title":"The effect of carbon dioxide on the intracellular pH and buffering power of snail neurones","volume":"255","author":"Thomas","year":"1976","journal-title":"J. Physiol."},{"key":"2023072918145169100_ThomasandMeech1982","doi-asserted-by":"crossref","first-page":"826","DOI":"10.1038\/299826a0","article-title":"Hydrogen ion currents and intracellular pH in depolarized voltage-clamped snail neurones","volume":"299","author":"Thomas","year":"1982","journal-title":"Nature."},{"key":"2023072918145169100_Wankeetal1979","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/S0006-3495(79)85251-0","article-title":"K+ conductance modified by a titratable group accessible to protons from the intracellular side of the squid axon membrane","volume":"26","author":"Wanke","year":"1979","journal-title":"Biophys. J."}],"container-title":["The Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/114\/6\/771\/1905536\/gp-7981.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/114\/6\/771\/1905536\/gp-7981.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,4]],"date-time":"2024-01-04T17:21:57Z","timestamp":1704388917000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/114\/6\/771\/45531\/Evidence-That-the-Product-of-the-Human-X-Linked"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,11,15]]},"references-count":43,"journal-issue":{"issue":"6","published-print":{"date-parts":[[1999,12,1]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.114.6.771","relation":{},"ISSN":["0022-1295","1540-7748"],"issn-type":[{"value":"0022-1295","type":"print"},{"value":"1540-7748","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,11,15]]}}}