Home LiteratureArticle Details
PMID: 26824658 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

LRRC8 Proteins Form Volume-Regulated Anion Channels that Sense Ionic Strength.

Cell ·Vol. 164 ·No. 3 ·2016-01-28 ·Pages 499-511

Syeda R, Qiu Z, Dubin AE, Murthy SE, Florendo MN, Mason DE, Mathur J, Cahalan SM, Peters EC, Montal M, Patapoutian A

Abstract

The volume-regulated anion channel (VRAC) is activated when a cell swells, and it plays a central role in maintaining cell volume in response to osmotic challenges. SWELL1 (LRRC8A) was recently identified as an essential component of VRAC. However, the identity of the pore-forming subunits of VRAC and how the channel is gated by cell swelling are unknown. Here, we show that SWELL1 and up to four other LRRC8 subunits assemble into heterogeneous complexes of ∼800 kDa. When reconstituted into bilayers, LRRC8 complexes are sufficient to form anion channels activated by osmolality gradients. In bilayers, as well as in cells, the single-channel conductance of the complexes depends on the LRRC8 composition. Finally, low ionic strength (Γ) in the absence of an osmotic gradient activates the complexes in bilayers. These data demonstrate that LRRC8 proteins together constitute the VRAC pore and that hypotonic stress can activate VRAC through a decrease in cytoplasmic Γ.

MeSH Terms
HeLa Cells Humans Ion Channels/chemistry,metabolism Lipid Bilayers/chemistry,metabolism Membrane Proteins/metabolism Multiprotein Complexes/chemistry,metabolism Osmosis
Chemicals
Ion Channels LRRC8A protein, human Lipid Bilayers Membrane Proteins Multiprotein Complexes
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Syeda Ruhma
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA.
Qiu Zhaozhu
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA; Genomics Institute of the Novartis Research Foundation (GNF), San Diego, CA 92121, USA.
Dubin Adrienne E
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: adubin@scripps.edu.
Murthy Swetha E
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA.
Florendo Maria N
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA.
Mason Daniel E
Genomics Institute of the Novartis Research Foundation (GNF), San Diego, CA 92121, USA.
Mathur Jayanti
Genomics Institute of the Novartis Research Foundation (GNF), San Diego, CA 92121, USA.
Cahalan Stuart M
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA.
Peters Eric C
Genomics Institute of the Novartis Research Foundation (GNF), San Diego, CA 92121, USA.
Montal Mauricio
Division of Biological Sciences, University of California, San Diego, La Jolla, CA 92093, USA.
Patapoutian Ardem
Department of Molecular and Cellular Neuroscience, Howard Hughes Medical Institute, Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: ardem@scripps.edu.
References (37)
37 references, click to expand
  1. Reduction of ionic strength activates single volume-regulated anion channels (VRAC) in endothelial cells.
    Pflugers Arch. 2000 Jan;439(3):315-20 PMID: 10650983
  2. Subunit composition of VRAC channels determines substrate specificity and cellular resistance to Pt-based anti-cancer drugs.
    EMBO J. 2015 Dec 14;34(24):2993-3008 PMID: 26530471
  3. The novel gene fad158, having a transmembrane domain and leucine-rich repeat, stimulates adipocyte differentiation.
    J Biol Chem. 2004 Aug 13;279(33):34840-8 PMID: 15184384
  4. Swelling-induced and depolarization-induced C1-channels in normal and cystic fibrosis epithelial cells.
    Am J Physiol. 1991 Oct;261(4 Pt 1):C658-74 PMID: 1656769
  5. Ion channels activated by swelling of Madin Darby canine kidney (MDCK) cells.
    J Membr Biol. 1992 Mar;126(2):109-14 PMID: 1317457
  6. Osmotic swelling activates intermediate-conductance Cl- channels in human intestinal epithelial cells.
    Jpn J Physiol. 1994;44(4):403-9 PMID: 7869602
  7. Characterization of the voltage-dependent properties of a volume-sensitive anion conductance.
    J Gen Physiol. 1995 May;105(5):661-76 PMID: 7544824
  8. Volume-activated Cl- channels.
    Gen Pharmacol. 1996 Oct;27(7):1131-40 PMID: 8981057
  9. Sensitivity of volume-sensitive Cl- conductance in human epithelial cells to extracellular nucleotides.
    Am J Physiol. 1996 Dec;271(6 Pt 1):C1872-8 PMID: 8997187
  10. Volume expansion-sensing outward-rectifier Cl- channel: fresh start to the molecular identity and volume sensor.
    Am J Physiol. 1997 Sep;273(3 Pt 1):C755-89 PMID: 9316396
  11. Activation of volume-regulated chloride currents by reduction of intracellular ionic strength in bovine endothelial cells.
    J Physiol. 1998 Jan 15;506 ( Pt 2):353-61 PMID: 9490864
  12. Intracellular ionic strength regulates the volume sensitivity of a swelling-activated anion channel.
    Am J Physiol. 1998 Aug;275(2 Pt 1):C416-22 PMID: 9688595
  13. Caveolin-1 modulates the activity of the volume-regulated chloride channel.
    J Physiol. 1999 Oct 1;520 Pt 1:113-9 PMID: 10517805
  14. Bacterial osmosensing: roles of membrane structure and electrostatics in lipid-protein and protein-protein interactions.
    Biochim Biophys Acta. 2004 Nov 3;1666(1-2):88-104 PMID: 15519310
  15. Salt bridges and gating in the COOH-terminal region of HCN2 and CNGA1 channels.
    J Gen Physiol. 2004 Dec;124(6):663-77 PMID: 15572346
  16. A phospholipid sensor controls mechanogating of the K+ channel TREK-1.
    EMBO J. 2005 Jan 12;24(1):44-53 PMID: 15577940
  17. Single-channel properties of volume-sensitive Cl- channel in ClC-3-deficient cardiomyocytes.
    Jpn J Physiol. 2005 Dec;55(6):379-83 PMID: 16441975
  18. A sensor for intracellular ionic strength.
    Proc Natl Acad Sci U S A. 2006 Jul 11;103(28):10624-9 PMID: 16815971
  19. Ion specificity and ionic strength dependence of the osmoregulatory ABC transporter OpuA.
    J Biol Chem. 2006 Oct 6;281(40):29830-9 PMID: 16844687
  20. Macromolecular crowding and confinement: biochemical, biophysical, and potential physiological consequences.
    Annu Rev Biophys. 2008;37:375-97 PMID: 18573087
  21. Screening blockers against a potassium channel with a droplet interface bilayer array.
    J Am Chem Soc. 2008 Nov 19;130(46):15543-8 PMID: 18950170
  22. Physiology of cell volume regulation in vertebrates.
    Physiol Rev. 2009 Jan;89(1):193-277 PMID: 19126758
  23. Rapid and improved reconstitution of bacterial mechanosensitive ion channel proteins MscS and MscL into liposomes using a modified sucrose method.
    FEBS Lett. 2009 Jan 22;583(2):407-12 PMID: 19111548
  24. Droplet interface bilayers.
    Mol Biosyst. 2008 Dec;4(12):1191-208 PMID: 19396383
  25. Mass estimation of native proteins by blue native electrophoresis: principles and practical hints.
    Mol Cell Proteomics. 2010 Oct;9(10):2149-61 PMID: 20173216
  26. Factor for adipocyte differentiation 158 gene disruption prevents the body weight gain and insulin resistance induced by a high-fat diet.
    Biol Pharm Bull. 2011;34(8):1257-63 PMID: 21804215
  27. LRRC8 proteins share a common ancestor with pannexins, and may form hexameric channels involved in cell-cell communication.
    Bioessays. 2012 Jul;34(7):551-60 PMID: 22532330
  28. Mechanosensitivity is mediated directly by the lipid membrane in TRAAK and TREK1 K+ channels.
    Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):3614-9 PMID: 24550493
  29. SWELL1, a plasma membrane protein, is an essential component of volume-regulated anion channel.
    Cell. 2014 Apr 10;157(2):447-58 PMID: 24725410
  30. Identification of LRRC8 heteromers as an essential component of the volume-regulated anion channel VRAC.
    Science. 2014 May 9;344(6184):634-8 PMID: 24790029
  31. Feeling the hidden mechanical forces in lipid bilayer is an original sense.
    Proc Natl Acad Sci U S A. 2014 Jun 3;111(22):7898-905 PMID: 24850861
  32. The protein synthesis inhibitor blasticidin s enters mammalian cells via leucine-rich repeat-containing protein 8D.
    J Biol Chem. 2014 Jun 13;289(24):17124-31 PMID: 24782309
  33. Characteristics and roles of the volume-sensitive outwardly rectifying (VSOR) anion channel in the central nervous system.
    Neuroscience. 2014 Sep 5;275:211-31 PMID: 24937753
  34. The multifaceted subunit interfaces of ionotropic glutamate receptors.
    J Physiol. 2015 Jan 1;593(1):73-81 PMID: 25556789
  35. The identification of a volume-regulated anion channel: an amazing Odyssey.
    Acta Physiol (Oxf). 2015 Apr;213(4):868-81 PMID: 25565132
  36. TMEM16, LRRC8A, bestrophin: chloride channels controlled by Ca(2+) and cell volume.
    Trends Biochem Sci. 2015 Sep;40(9):535-43 PMID: 26254230
  37. LRRC8 involved in B cell development belongs to a novel family of leucine-rich repeat proteins.
    FEBS Lett. 2004 Apr 23;564(1-2):147-52 PMID: 15094057
Article Info
Journal
Cell
Abbr.
Cell
ISSN
1097-4172
DOI
10.1016/j.cell.2015.12.031
Email
adubin@scripps.edu
ardem@scripps.edu
Published
2016-01-28
Pages
499-511
Language
English
Region
United States
NLM ID
0413066
PMCID
PMC4733249
Subset
IM
Grants
NINDS NIH HHS · R01 NS083174 · United States
NINDS NIH HHS · NS083174 · United States
NIGMS NIH HHS · R01 GM049711 · United States
Howard Hughes Medical Institute · United States
NIGMS NIH HHS · GM49711 · United States
External Links
PubMed source DOI: 10.1016/j.cell.2015.12.031 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2