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Cited by 35Ion channel regulation and functionIon Transport and Channel RegulationIon Channels and Receptors

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Cryo-EM structures of an LRRC8 chimera with native functional properties reveal heptameric assembly

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Takahashi, Hirohide, Yamada, Toshiki, Denton, Jerod S, Strange, Kevin, Karakas, Erkan·2023·eLife
DOI 10.7554/elife.82431PMC PMC10049205

Published version of a preprint

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Abstract

Volume-regulated anion channels (VRACs) mediate volume regulatory Cl - and organic solute efflux from vertebrate cells. VRACs are heteromeric assemblies of LRRC8A-E proteins with unknown stoichiometries. Homomeric LRRC8A and LRRC8D channels have a small pore, hexameric structure. However, these channels are either non-functional or exhibit abnormal regulation and pharmacology, limiting their utility for structure-function analyses. We circumvented these limitations by developing novel homomeric LRRC8 chimeric channels with functional properties consistent with those of native VRAC/LRRC8 channels. We demonstrate here that the LRRC8C-LRRC8A(IL1 25 ) chimera comprising LRRC8C and 25 amino acids unique to the first intracellular loop (IL1) of LRRC8A has a heptameric structure like that of homologous pannexin channels. Unlike homomeric LRRC8A and LRRC8D channels, heptameric LRRC8C-LRRC8A(IL1 25 ) channels have a large-diameter pore similar to that estimated for native VRACs, exhibit normal DCPIB pharmacology, and have higher permeability to large organic anions. Lipid-like densities are located between LRRC8C-LRRC8A(IL1 25 ) subunits and occlude the channel pore. Our findings provide new insights into VRAC/LRRC8 channel structure and suggest that lipids may play important roles in channel gating and regulation.

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Metadata

Authors
Takahashi, H., Yamada, T., Denton, J. S., Strange, K., & Karakas, E.
Affiliations
  • Vanderbilt University (US)
  • Vanderbilt University Medical Center (US)
  • Department of Molecular Physiology and Biophysics, School of Medicine, Vanderbilt University
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2023-03-10
Volume
12
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.82431
PMC
PMC10049205
OpenAlex
W4323812162
Cited by
35
References
2
Citations by year
2026: 11 · 2025: 8 · 2024: 8 · 2023: 7
FWCI
3.02
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Institute of Diabetes and Digestive and Kidney Diseases (DK51610)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
Keywords
volume-regulated anion channel, LRRC8A, chimeric channels, channel structure, DCPIB, channel gating, pannexin channels, homomeric channels
MeSH terms
Anions, Biological Transport, Membrane Proteins, Cryoelectron Microscopy

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