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PMID: 23630341 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

TMEM16F (Anoctamin 6), an anion channel of delayed Ca(2+) activation.

The Journal of general physiology ·Vol. 141 ·No. 5 ·2013-05-00 ·Pages 585-600

Grubb S, Poulsen KA, Juul CA, Kyed T, Klausen TK, Larsen EH, Hoffmann EK

Abstract

Members of the TMEM16 (Anoctamin) family of membrane proteins have been shown to be essential constituents of the Ca(2+)-activated Cl(-) channel (CaCC) in many cell types. In this study, we have investigated the electrophysiological properties of mouse TMEM16F. Heterologous expression of TMEM16F in HEK293 cells resulted in plasma membrane localization and an outwardly rectifying ICl,Ca that was activated with a delay of several minutes. Furthermore, a significant Na(+) current was activated, and the two permeabilities were correlated according to PNa = 0.3 PCl. The current showed an EC50 of 100 µM intracellular free Ca(2+) concentration and an Eisenman type 1 anion selectivity sequence of PSCN > PI > PBr > PCl > PAsp. The mTMEM16F-associated ICl,Ca was abolished in one mutant of the putative pore region (R592E) but retained in two other mutants (K616E and R636E). The mutant K616E had a lower relative permeability to iodide, and the mutant R636E had an altered anion selectivity sequence (PSCN = PI = PBr = PCl > PAsp). Our data provide evidence that TMEM16F constitutes a Ca(2+)-activated anion channel or a pore-forming subunit of an anion channel with properties distinct from TMEM16A.

MeSH Terms
Animals Anions/metabolism Anoctamins Calcium/metabolism Cell Membrane/metabolism,physiology Cells, Cultured Chloride Channels/metabolism HEK293 Cells Humans Ion Transport/physiology Membrane Potentials/physiology Membrane Proteins/metabolism Mice Permeability Phospholipid Transfer Proteins/metabolism Sodium/metabolism
Chemicals
ANO6 protein, mouse Anions Anoctamins Chloride Channels Membrane Proteins Phospholipid Transfer Proteins Sodium Calcium
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Grubb Søren
Department of Biology, University of Copenhagen, DK-2100 Copenhagen Ø, Denmark.
Poulsen Kristian A
Juul Christian Ammitzbøll
Kyed Tania
Klausen Thomas K
Larsen Erik Hviid
Hoffmann Else K
References (59)
59 references, click to expand
  1. Expression and function of epithelial anoctamins.
    J Biol Chem. 2010 Mar 5;285(10):7838-45 PMID: 20056604
  2. ANOs 3-7 in the anoctamin/Tmem16 Cl- channel family are intracellular proteins.
    Am J Physiol Cell Physiol. 2012 Feb 1;302(3):C482-93 PMID: 22075693
  3. Molecular structure and physiological function of chloride channels.
    Physiol Rev. 2002 Apr;82(2):503-68 PMID: 11917096
  4. Expression and function of epithelial anoctamins.
    Exp Physiol. 2012 Feb;97(2):184-92 PMID: 21908539
  5. Anoctamin 6 is an essential component of the outwardly rectifying chloride channel.
    Proc Natl Acad Sci U S A. 2011 Nov 1;108(44):18168-72 PMID: 22006324
  6. The transmembrane protein TMEM16A is required for normal development of the murine trachea.
    Dev Biol. 2008 Sep 1;321(1):141-9 PMID: 18585372
  7. TMEM16A confers receptor-activated calcium-dependent chloride conductance.
    Nature. 2008 Oct 30;455(7217):1210-5 PMID: 18724360
  8. The novel marker, DOG1, is expressed ubiquitously in gastrointestinal stromal tumors irrespective of KIT or PDGFRA mutation status.
    Am J Pathol. 2004 Jul;165(1):107-13 PMID: 15215166
  9. Expression of anoctamin 1/TMEM16A by interstitial cells of Cajal is fundamental for slow wave activity in gastrointestinal muscles.
    J Physiol. 2009 Oct 15;587(Pt 20):4887-904 PMID: 19687122
  10. Compound heterozygosity for 2 novel TMEM16F mutations in a patient with Scott syndrome.
    Blood. 2011 Apr 21;117(16):4399-400 PMID: 21511967
  11. Topology of NGEP, a prostate-specific cell:cell junction protein widely expressed in many cancers of different grade level.
    Cancer Res. 2008 Aug 1;68(15):6306-12 PMID: 18676855
  12. Expanding the range of free calcium regulation in biological solutions.
    Anal Biochem. 2005 Dec 15;347(2):303-15 PMID: 16289079
  13. Molecular cloning and characterization of the murine gnathodiaphyseal dysplasia gene GDD1.
    Biochem Biophys Res Commun. 2005 Jun 17;331(4):1099-106 PMID: 15882990
  14. Kinetic and pharmacological properties of the calcium-activated chloride-current in macrovascular endothelial cells.
    Cell Calcium. 1997 Jul;22(1):53-63 PMID: 9232352
  15. Advances in our understanding of the molecular basis of disorders of platelet function.
    J Thromb Haemost. 2011 Jul;9 Suppl 1:76-91 PMID: 21781244
  16. Transmembrane protein 16A (TMEM16A) is a Ca2+-regulated Cl- secretory channel in mouse airways.
    J Biol Chem. 2009 May 29;284(22):14875-80 PMID: 19363029
  17. Calcium-dependent phospholipid scrambling by TMEM16F.
    Nature. 2010 Dec 9;468(7325):834-8 PMID: 21107324
  18. Calcium-activated chloride current contributes to action potential alternations in left ventricular hypertrophy rabbit.
    Am J Physiol Heart Circ Physiol. 2008 Jul;295(1):H97-H104 PMID: 18441200
  19. TMEM16 proteins produce volume-regulated chloride currents that are reduced in mice lacking TMEM16A.
    J Biol Chem. 2009 Oct 16;284(42):28571-8 PMID: 19654323
  20. P2 receptor-mediated signal transduction in Ehrlich ascites tumor cells.
    Biochim Biophys Acta. 1998 Sep 23;1374(1-2):94-106 PMID: 9814856
  21. TMEM16F is a component of a Ca2+-activated Cl- channel but not a volume-sensitive outwardly rectifying Cl- channel.
    Am J Physiol Cell Physiol. 2013 Apr 15;304(8):C748-59 PMID: 23426967
  22. Loss of TMEM16A causes a defect in epithelial Ca2+-dependent chloride transport.
    J Biol Chem. 2009 Oct 16;284(42):28698-703 PMID: 19679661
  23. Calcium-activated chloride channels: (un)known, (un)loved?
    Proc Am Thorac Soc. 2004;1(1):22-7 PMID: 16113407
  24. TMEM16A, a membrane protein associated with calcium-dependent chloride channel activity.
    Science. 2008 Oct 24;322(5901):590-4 PMID: 18772398
  25. Evolution and functional divergence of the anoctamin family of membrane proteins.
    BMC Evol Biol. 2010 Oct 21;10:319 PMID: 20964844
  26. TMEM16B, a novel protein with calcium-dependent chloride channel activity, associates with a presynaptic protein complex in photoreceptor terminals.
    J Neurosci. 2009 May 27;29(21):6809-18 PMID: 19474308
  27. TMEM16A(a)/anoctamin-1 shares a homodimeric architecture with CLC chloride channels.
    Mol Cell Proteomics. 2011 Feb;10(2):M110.004697 PMID: 20974900
  28. Reconciling the Krogh and Ussing interpretations of epithelial chloride transport - presenting a novel hypothesis for the physiological significance of the passive cellular chloride uptake.
    Acta Physiol (Oxf). 2011 Jul;202(3):435-64 PMID: 21288306
  29. Bestrophin and TMEM16-Ca(2+) activated Cl(-) channels with different functions.
    Cell Calcium. 2009 Oct;46(4):233-41 PMID: 19783045
  30. Expression of TMEM16 paralogs during murine embryogenesis.
    Dev Dyn. 2008 Sep;237(9):2566-74 PMID: 18729231
  31. Expression patterns of the Tmem16 gene family during cephalic development in the mouse.
    Gene Expr Patterns. 2009 Mar;9(3):178-91 PMID: 19059364
  32. Calcium-activated chloride channels in the retina.
    Channels (Austin). 2008 Jul-Aug;2(4):252-60 PMID: 18769137
  33. Calcium, the two-faced messenger of olfactory transduction and adaptation.
    Curr Opin Neurobiol. 2003 Aug;13(4):469-75 PMID: 12965295
  34. Anoctamin/TMEM16 family members are Ca2+-activated Cl- channels.
    J Physiol. 2009 May 15;587(Pt 10):2127-39 PMID: 19015192
  35. A transient calcium-dependent chloride current in the immature Xenopus oocyte.
    J Physiol. 1983 Sep;342:309-25 PMID: 6313909
  36. Physiological roles and diseases of Tmem16/Anoctamin proteins: are they all chloride channels?
    Acta Pharmacol Sin. 2011 Jun;32(6):685-92 PMID: 21642943
  37. NGEP, a gene encoding a membrane protein detected only in prostate cancer and normal prostate.
    Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):3059-64 PMID: 14981236
  38. GDD1 is identical to TMEM16E, a member of the TMEM16 family.
    Am J Hum Genet. 2004 Nov;75(5):927-8; author reply 928-9 PMID: 15457408
  39. Characterization of the oligomeric structure of the Ca(2+)-activated Cl- channel Ano1/TMEM16A.
    J Biol Chem. 2011 Jan 14;286(2):1381-8 PMID: 21056985
  40. Chloride channels as drug targets.
    Nat Rev Drug Discov. 2009 Feb;8(2):153-71 PMID: 19153558
  41. Expression cloning of TMEM16A as a calcium-activated chloride channel subunit.
    Cell. 2008 Sep 19;134(6):1019-29 PMID: 18805094
  42. Separate swelling- and Ca2+-activated anion currents in Ehrlich ascites tumor cells.
    J Membr Biol. 1998 May 15;163(2):97-110 PMID: 9592074
  43. Physiology of cell volume regulation in vertebrates.
    Physiol Rev. 2009 Jan;89(1):193-277 PMID: 19126758
  44. Amazing chloride channels: an overview.
    Acta Physiol Scand. 2003 Feb;177(2):119-47 PMID: 12558550
  45. Bimodal control of a Ca(2+)-activated Cl(-) channel by different Ca(2+) signals.
    J Gen Physiol. 2000 Jan;115(1):59-80 PMID: 10613919
  46. TMEM16B induces chloride currents activated by calcium in mammalian cells.
    Pflugers Arch. 2009 Oct;458(6):1023-38 PMID: 19475416
  47. The anoctamin family: TMEM16A and TMEM16B as calcium-activated chloride channels.
    Exp Physiol. 2012 Feb;97(2):177-83 PMID: 21984732
  48. Explaining calcium-dependent gating of anoctamin-1 chloride channels requires a revised topology.
    Circ Res. 2012 Mar 30;110(7):990-9 PMID: 22394518
  49. Anoctamin 2/TMEM16B: a calcium-activated chloride channel in olfactory transduction.
    Exp Physiol. 2012 Feb;97(2):193-9 PMID: 21890523
  50. Tmem16b is specifically expressed in the cilia of olfactory sensory neurons.
    Chem Senses. 2010 Mar;35(3):239-45 PMID: 20100788
  51. Mechanism of the inhibition of Ca2+-activated Cl- currents by phosphorylation in pulmonary arterial smooth muscle cells.
    J Gen Physiol. 2006 Jul;128(1):73-87 PMID: 16801382
  52. Calcium-dependent chloride conductance in epithelia: is there a contribution by Bestrophin?
    Pflugers Arch. 2007 Sep;454(6):879-89 PMID: 17361457
  53. TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation.
    Cell. 2012 Sep 28;151(1):111-22 PMID: 23021219
  54. Calcium-activated chloride channels.
    Annu Rev Physiol. 2005;67:719-58 PMID: 15709976
  55. Axotomy-induced expression of calcium-activated chloride current in subpopulations of mouse dorsal root ganglion neurons.
    J Neurophysiol. 2003 Dec;90(6):3764-73 PMID: 12944538
  56. Properties of acetylcholine receptors translated by cat muscle mRNA in Xenopus oocytes.
    EMBO J. 1982;1(11):1307-12 PMID: 6144542
  57. Recessive mutations in the putative calcium-activated chloride channel Anoctamin 5 cause proximal LGMD2L and distal MMD3 muscular dystrophies.
    Am J Hum Genet. 2010 Feb 12;86(2):213-21 PMID: 20096397
  58. Voltage-activated and calcium-activated currents studied in solitary rod inner segments from the salamander retina.
    J Physiol. 1982 Oct;331:253-84 PMID: 7153904
  59. Cell cycle-dependent activity of the volume- and Ca2+-activated anion currents in Ehrlich lettre ascites cells.
    J Cell Physiol. 2007 Mar;210(3):831-42 PMID: 17111356
Article Info
Journal
The Journal of general physiology
Abbr.
J Gen Physiol
ISSN
1540-7748
DOI
10.1085/jgp.201210861
Published
2013-05-00
Pages
585-600
Language
English
Region
United States
NLM ID
2985110R
PMCID
PMC3639583
Subset
IM
External Links
PubMed source DOI: 10.1085/jgp.201210861 PMC Full Text View journal details
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