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Structure paper

TitleCryo-EM structures of the DCPIB-inhibited volume-regulated anion channel LRRC8A in lipid nanodiscs.
Journal, issue, pagesElife, Vol. 8, Year 2019
Publish dateFeb 18, 2019
AuthorsDavid M Kern / SeCheol Oh / Richard K Hite / Stephen G Brohawn /
PubMed AbstractHypoosmotic conditions activate volume-regulated anion channels in vertebrate cells. These channels are formed by leucine-rich repeat-containing protein 8 (LRRC8) family members and contain LRRC8A in ...Hypoosmotic conditions activate volume-regulated anion channels in vertebrate cells. These channels are formed by leucine-rich repeat-containing protein 8 (LRRC8) family members and contain LRRC8A in homo- or hetero-hexameric assemblies. Here, we present single-particle cryo-electron microscopy structures of LRRC8A in complex with the inhibitor DCPIB reconstituted in lipid nanodiscs. DCPIB plugs the channel like a cork in a bottle - binding in the extracellular selectivity filter and sterically occluding ion conduction. Constricted and expanded structures reveal coupled dilation of cytoplasmic LRRs and the channel pore, suggesting a mechanism for channel gating by internal stimuli. Conformational and symmetry differences between LRRC8A structures determined in detergent micelles and lipid bilayers related to reorganization of intersubunit lipid binding sites demonstrate a critical role for the membrane in determining channel structure. These results provide insight into LRRC8 gating and inhibition and the role of lipids in the structure of an ionic-strength sensing ion channel.
External linksElife / PubMed:30775971 / PubMed Central
MethodsEM (single particle)
Resolution3.21 - 4.18 Å
Structure data

0562

6nzw

EMDB-0562, PDB-6nzw:
LRRC8A-DCPIB in MSP1E3D1 nanodisc constricted state
Method: EM (single particle) / Resolution: 3.21 Å

0563

6nzz

EMDB-0563, PDB-6nzz:
LRRC8A-DCPIB in MSP1E3D1 nanodisc expanded state
Method: EM (single particle) / Resolution: 3.32 Å

0564

6o00

EMDB-0564, PDB-6o00:
apo-LRRC8A in MSP2N2 nanodisc constricted state
Method: EM (single particle) / Resolution: 4.18 Å

Chemicals

ChemComp-POV:
(2S)-3-(hexadecanoyloxy)-2-[(9Z)-octadec-9-enoyloxy]propyl 2-(trimethylammonio)ethyl phosphate / phospholipid*YM / POPC

ChemComp-L9Y:
4-{[(2S)-2-butyl-6,7-dichloro-2-cyclopentyl-1-oxo-2,3-dihydro-1H-inden-5-yl]oxy}butanoic acid

Source
  • mus musculus (house mouse)
KeywordsMEMBRANE PROTEIN / Ion channel / volume-regulation

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