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Open data
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Basic information
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Title | Structure of LRRC8C-LRRC8A(IL125) Chimera, Class 2 | |||||||||
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Function / homology | ![]() pre-B cell differentiation / Miscellaneous transport and binding events / volume-sensitive anion channel activity / taurine transmembrane transport / aspartate transmembrane transport / cyclic-GMP-AMP transmembrane transporter activity / cyclic-GMP-AMP transmembrane import across plasma membrane / monoatomic anion transmembrane transport / cellular response to osmotic stress / cell volume homeostasis ...pre-B cell differentiation / Miscellaneous transport and binding events / volume-sensitive anion channel activity / taurine transmembrane transport / aspartate transmembrane transport / cyclic-GMP-AMP transmembrane transporter activity / cyclic-GMP-AMP transmembrane import across plasma membrane / monoatomic anion transmembrane transport / cellular response to osmotic stress / cell volume homeostasis / protein hexamerization / monoatomic anion transport / response to osmotic stress / monoatomic ion channel complex / fat cell differentiation / intracellular glucose homeostasis / positive regulation of myoblast differentiation / chloride transmembrane transport / positive regulation of insulin secretion / spermatogenesis / lysosomal membrane / endoplasmic reticulum membrane / cell surface / identical protein binding / membrane / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
![]() | Takahashi H / Yamada T / Denton JS / Strange K / Karakas E | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structures of an LRRC8 chimera with native functional properties reveal heptameric assembly. Authors: Hirohide Takahashi / Toshiki Yamada / Jerod S Denton / Kevin Strange / Erkan Karakas / ![]() 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 ...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) 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) 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) 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. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 168.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18.7 KB 18.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.9 KB | Display | ![]() |
Images | ![]() | 49.3 KB | ||
Others | ![]() ![]() ![]() | 89 MB 165.3 MB 165.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8dxoMC ![]() 8dxnC ![]() 8dxpC ![]() 8dxqC ![]() 8dxrC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 1.1 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Unsharpened map.
File | emd_27771_additional_1.map | ||||||||||||
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Annotation | Unsharpened map. | ||||||||||||
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Density Histograms |
-Half map: #2
File | emd_27771_half_map_1.map | ||||||||||||
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Density Histograms |
-Half map: #1
File | emd_27771_half_map_2.map | ||||||||||||
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Density Histograms |
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Sample components
-Entire : LRRC8C-LRRC8A(IL125) chimera
Entire | Name: LRRC8C-LRRC8A(IL125) chimera |
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Components |
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-Supramolecule #1: LRRC8C-LRRC8A(IL125) chimera
Supramolecule | Name: LRRC8C-LRRC8A(IL125) chimera / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: all / Details: Heptameric LRRC8C-LRRC8A(IL125) chimera. |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 700 KDa |
-Macromolecule #1: Volume-regulated anion channel subunit LRRC8C,Volume-regulated an...
Macromolecule | Name: Volume-regulated anion channel subunit LRRC8C,Volume-regulated anion channel subunit LRRC8A type: protein_or_peptide / ID: 1 Details: LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where ...Details: LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag. Number of copies: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 94.747695 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MIPVTEFRQF SEQQPAFRVL KPWWDVFTDY LSVAMLMIGV FGCTLQVMQD KIICLPKRVQ PAQNHSSLSN VSQAVASTTP LPPPKPSPA NPITVEMKGL KTDLDLQQYS FINQMCYERA LHWYAKYFPY LVLIHTLVFM LCSNFWFKFP GSSSKIEHFI S ILGKCFDS ...String: MIPVTEFRQF SEQQPAFRVL KPWWDVFTDY LSVAMLMIGV FGCTLQVMQD KIICLPKRVQ PAQNHSSLSN VSQAVASTTP LPPPKPSPA NPITVEMKGL KTDLDLQQYS FINQMCYERA LHWYAKYFPY LVLIHTLVFM LCSNFWFKFP GSSSKIEHFI S ILGKCFDS PWTTRALSEV SGEDSDPKPA FSKMNGSMDK KSSTVSEDVE GSLVNSQSLK SIPEKFVVDK STAGALDKKE GE QAKALFE KVKKFRLHVE EGDILYAMYV RQTVLKVIKF LIIIAYNSAL VSKVQFTVDC NVDIQDMTGY KNFSCNHTMA HLF SKLSFC YLCFVSIYGL TCLYTLYWLF YRSLREYSFE YVRQETGIDD IPDVKNDFAF MLHMIDQYDP LYSKRFAVFL SEVS ENKLK QLNLNNEWTP DKLRQKLQTN AHNRLELPLI MLSGLPDTVF EITELQSLKL EIIKNVMIPA TIAQLDNLQE LSLHQ CSVK IHSAALSFLK ENLKVLSVKF DDMRELPPWM YGLRNLEELY LVGSLSHDIS RNVTLESLRD LKSLKILSIK SNVSKI PQA VVDVSSHLQK MCIHNDGTKL VMLNNLKKMT NLTELELVHC DLERIPHAVF SLLSLQELDL KENNLKSIEE IVSFQHL RK LTVLKLWHNS ITYIPEHIKK LTSLERLSFS HNKIEVLPSH LFLCNKIRYL DLSYNDIRFI PPEIGVLQSL QYFSITCN V ESLPDELYFC KKLKTLKIGK NSLSVLSPKI GNLLFLSYLD VKGNHFEILP PELGDCRALK RAGLVVEDAL FETLPSDVR EQMKTEENLY FQGAAAGDYK DDDDK |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 3.0 mg/mL | ||||||||||||
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Buffer | pH: 8 Component:
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Grid | Model: UltrAuFoil R1.2/1.3 / Mesh: 300 / Support film - Material: GOLD / Support film - topology: HOLEY | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 281 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 3198 / Average electron dose: 54.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
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Output model | ![]() PDB-8dxo: |