Voltage changes across the cell membrane control the gating of many cation-selective ion channels. Conserved from bacteria to humans1, the voltage-gated-ligand superfamily of ion channels are encoded as polypeptide chains of six transmembrane-spanning segments (S1–S6). S1–S4 functions as a self-contained voltage-sensing domain (VSD), in essence a positively charged lever that moves in response to voltage changes. The VSD ‘ligand’ transmits force via a linker to the S5–S6 pore domain ‘receptor’2, thereby opening or closing the channel. The ascidian VSD protein Ci-VSP gates a phosphatase activity rather than a channel pore, indicating that VSDs function independently of ion channels3. Here we describe a mammalian VSD protein (HV1) that lacks a discernible pore domain but is sufficient for expression of a voltage-sensitive proton-selective ion channel activity. Hv1 currents are activated at depolarizing voltages, sensitive to the transmembrane pH gradient, H+-selective, and Zn2+-sensitive. Mutagenesis of Hv1 identified three arginine residues in S4 that regulate channel gating and two histidine residues that are required for extracellular inhibition of Hv1 by Zn2+. Hv1 is expressed in immune tissues and manifests the characteristic properties of native proton conductances (). In phagocytic leukocytes4, are required to support the oxidative burst that underlies microbial killing by the innate immune system4,5. The data presented here identify Hv1 as a long-sought voltage-gated H+ channel and establish Hv1 as the founding member of a family of mammalian VSD proteins.
Your institute does not have access to this article
Open Access articles citing this article.
Nature Cell Biology Open Access 19 May 2022
The voltage-gated proton channel Hv1 promotes microglia-astrocyte communication and neuropathic pain after peripheral nerve injury
Molecular Brain Open Access 28 June 2021
Direct activation of the proton channel by albumin leads to human sperm capacitation and sustained release of inflammatory mediators by neutrophils
Nature Communications Open Access 22 June 2021
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Yu, F. H. & Catterall, W. A. The VGL-chanome: a protein superfamily specialized for electrical signaling and ionic homeostasis. Sci. STKE 2004, re15 (2004)
Long, S. B., Campbell, E. B. & Mackinnon, R. Crystal structure of a mammalian voltage-dependent Shaker family K+ channel. Science 309, 897–903 (2005)
Murata, Y., Iwasaki, H., Sasaki, M., Inaba, K. & Okamura, Y. Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. Nature 435, 1239–1243 (2005)
DeCoursey, T. E. Voltage-gated proton channels and other proton transfer pathways. Physiol. Rev. 83, 475–579 (2003)
DeCoursey, T. E., Morgan, D. & Cherny, V. V. The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels. Nature 422, 531–534 (2003)
Goldberg, J. et al. Three-dimensional structure of the catalytic subunit of protein serine/threonine phosphatase-1. Nature 376, 745–753 (1995)
Schilling, T., Gratopp, A., DeCoursey, T. E. & Eder, C. Voltage-activated proton currents in human lymphocytes. J. Physiol. (Lond.) 545, 93–105 (2002)
DeCoursey, T. E., Cherny, V. V., DeCoursey, A. G., Xu, W. & Thomas, L. L. Interactions between NADPH oxidase-related proton and electron currents in human eosinophils. J. Physiol. (Lond.) 535, 767–781 (2001)
Thomas, R. C. & Meech, R. W. Hydrogen ion currents and intracellular pH in depolarized voltage-clamped snail neurones. Nature 299, 826–828 (1982)
Henderson, L. M., Chappell, J. B. & Jones, O. T. Superoxide generation by the electrogenic NADPH oxidase of human neutrophils is limited by the movement of a compensating charge. Biochem. J. 255, 285–290 (1988)
Rada, B. K., Geiszt, M., Kaldi, K., Timar, C. & Ligeti, E. Dual role of phagocytic NADPH oxidase in bacterial killing. Blood 104, 2947–2953 (2004)
Babior, B. M. NADPH oxidase. Curr. Opin. Immunol. 16, 42–47 (2004)
Smith, R. M. & Curnutte, J. T. Molecular basis of chronic granulomatous disease. Blood 77, 673–686 (1991)
DeCoursey, T. E. & Cherny, V. V. Temperature dependence of voltage-gated H+ currents in human neutrophils, rat alveolar epithelial cells, and mammalian phagocytes. J. Gen. Physiol. 112, 503–522 (1998)
Cherny, V. V. & DeCoursey, T. E. pH-dependent inhibition of voltage-gated H+ currents in rat alveolar epithelial cells by Zn2+ and other divalent cations. J. Gen. Physiol. 114, 819–838 (1999)
Eder, C. & DeCoursey, T. E. Voltage-gated proton channels in microglia. Prog. Neurobiol. 64, 277–305 (2001)
Cherny, V. V., Henderson, L. M. & DeCoursey, T. E. Proton and chloride currents in Chinese hamster ovary cells. Membr. Cell Biol. 11, 337–347 (1997)
Morgan, D., Cherny, V. V., Price, M. O., Dinauer, M. C. & DeCoursey, T. E. Absence of proton channels in COS-7 cells expressing functional NADPH oxidase components. J. Gen. Physiol. 119, 571–580 (2002)
Bezanilla, F. The voltage sensor in voltage-dependent ion channels. Physiol. Rev. 80, 555–592 (2000)
Tombola, F., Pathak, M. M. & Isacoff, E. Y. How far will you go to sense voltage? Neuron 48, 719–725 (2005)
Starace, D. M. & Bezanilla, F. A proton pore in a potassium channel voltage sensor reveals a focused electric field. Nature 427, 548–553 (2004)
Pomes, R. & Roux, B. Molecular mechanism of H+ conduction in the single-file water chain of the gramicidin channel. Biophys. J. 82, 2304–2316 (2002)
DeCoursey, T. E., Morgan, D. & Cherny, V. V. The gp91phox component of NADPH oxidase is not a voltage-gated proton channel. J. Gen. Physiol. 120, 773–779 (2002)
Henderson, L. M. & Meech, R. W. Evidence that the product of the human X-linked CGD gene, gp91-phox, is a voltage-gated H+ pathway. J. Gen. Physiol. 114, 771–786 (1999)
Bokoch, G. M. Regulation of innate immunity by Rho GTPases. Trends Cell Biol. 15, 163–171 (2005)
DeCoursey, T. E., Cherny, V. V., Morgan, D., Katz, B. Z. & Dinauer, M. C. The gp91phox component of NADPH oxidase is not the voltage-gated proton channel in phagocytes, but it helps. J. Biol. Chem. 276, 36063–36066 (2001)
Price, M. O. et al. Creation of a genetic system for analysis of the phagocyte respiratory burst: high-level reconstitution of the NADPH oxidase in a nonhematopoietic system. Blood 99, 2653–2661 (2002)
Bokoch, G. M. & Knaus, U. G. NADPH oxidases: not just for leukocytes anymore! Trends Biochem. Sci. 28, 502–508 (2003)
Qu, A. Y., Nanda, A., Curnutte, J. T. & Grinstein, S. Development of a H+-selective conductance during granulocytic differentiation of HL-60 cells. Am. J. Physiol. 266, C1263–C1270 (1994)
Sasaki, M., Takagi, M. & Okamura, Y. A novel protein with a voltage sensor domain is a voltage-gated proton channel. Science (in the press)
We thank T. DeCoursey, C. Miller, R. MacKinnon and P. Bezanilla for comments on the manuscript, and K.-H. Lee for technical assistance. This work was supported by the Sandler Program for Asthma Research and the Howard Hughes Medical Institute.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
This file contains text to accompany the Supplementary Figures. (DOC 56 kb)
Hv1 amino acid sequence and expression profile. (PDF 1523 kb)
a, Human tissue Western blot probed with 4234 antibody (5 µg/ml) demonstrates expression of native Hv1 protein (~32 kDa) in immune tissues. b, Western blot of total cell lysates prepared from Jurkat (lane 1) or HEK-293T cells transfected with the indicated cDNA (lanes 3-5). c, in HL-60 cells, Hv1 protein appeared to be increased when cells were cultured in the presence of 1.3% DMSO (lane 2). (PDF 1536 kb)
a, Hv1-like currents were not detectable in HM1 cells. b, Native GvH+ in a DMSO-differentiated HL-60 cell. c, Temperature dependence of Hv1 currents. d, Monoexponential fits of τACT from the same cell as shown in panel c illustrate strong temperature-dependence of Hv1 kinetics. e, R205A currents. f, R208A currents. g, R211A currents. (PDF 2674 kb)
About this article
Cite this article
Ramsey, I., Moran, M., Chong, J. et al. A voltage-gated proton-selective channel lacking the pore domain. Nature 440, 1213–1216 (2006). https://doi.org/10.1038/nature04700
Inhibiting Hv1 channel in peripheral sensory neurons attenuates chronic inflammatory pain and opioid side effects
Cell Research (2022)
Cell Research (2022)
Nature Cell Biology (2022)
Journal of Physiology and Biochemistry (2022)
Intracellular pH regulation: characterization and functional investigation of H+ transporters in Stylophora pistillata
BMC Molecular and Cell Biology (2021)