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Open data
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Basic information
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Title | LRRC8A(T48D):C conformation 2 top focus | |||||||||
![]() | LRRC8A(T48D):C conformation 1 LRR focus 1 | |||||||||
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![]() | ion channel / volume-regulation / MEMBRANE PROTEIN | |||||||||
Function / homology | ![]() Miscellaneous transport and binding events / pre-B cell differentiation / 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 ...Miscellaneous transport and binding events / pre-B cell differentiation / 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 / electron transport chain / positive regulation of insulin secretion / spermatogenesis / periplasmic space / electron transfer activity / iron ion binding / lysosomal membrane / heme binding / endoplasmic reticulum membrane / cell surface / endoplasmic reticulum / identical protein binding / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.2 Å | |||||||||
![]() | Kern DM / Brohawn SG | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis for assembly and lipid-mediated gating of LRRC8A:C volume-regulated anion channels. Authors: David M Kern / Julia Bleier / Somnath Mukherjee / Jennifer M Hill / Anthony A Kossiakoff / Ehud Y Isacoff / Stephen G Brohawn / ![]() Abstract: Leucine-rich repeat-containing protein 8 (LRRC8) family members form volume-regulated anion channels activated by hypoosmotic cell swelling. LRRC8 channels are ubiquitously expressed in vertebrate ...Leucine-rich repeat-containing protein 8 (LRRC8) family members form volume-regulated anion channels activated by hypoosmotic cell swelling. LRRC8 channels are ubiquitously expressed in vertebrate cells as heteromeric assemblies of LRRC8A (SWELL1) and LRRC8B-E subunits. Channels of different subunit composition have distinct properties that explain the functional diversity of LRRC8 currents across cell types. However, the basis for heteromeric LRRC8 channel assembly and function is unknown. Here we leverage a fiducial-tagging strategy to determine single-particle cryo-EM structures of heterohexameric LRRC8A:C channels in multiple conformations. Compared to homomers, LRRC8A:C channels show pronounced differences in architecture due to heterotypic LRR interactions that displace subunits away from the conduction axis and poise the channel for activation. Structures and functional studies further reveal that lipids embedded in the channel pore block ion conduction in the closed state. These results provide insight into determinants for heteromeric LRRC8 channel assembly, activity and gating by lipids. | |||||||||
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 259.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19.6 KB 19.6 KB | Display Display | ![]() |
Images | ![]() | 54.9 KB | ||
Filedesc metadata | ![]() | 6.4 KB | ||
Others | ![]() ![]() ![]() | 136.9 MB 254.8 MB 254.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8f7dMC ![]() 8dr8C ![]() 8draC ![]() 8dreC ![]() 8drkC ![]() 8drnC ![]() 8droC ![]() 8drqC ![]() 8ds3C ![]() 8ds9C ![]() 8dsaC ![]() 8f74C ![]() 8f75C ![]() 8f77C ![]() 8f79C ![]() 8f7bC ![]() 8f7eC ![]() 8f7jC C: citing same article ( M: atomic model generated by this map |
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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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Annotation | LRRC8A(T48D):C conformation 1 LRR focus 1 | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.048 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: LRRC8A(T48D):C conformation 1 LRR focus 1 unsharpened
File | emd_28903_additional_1.map | ||||||||||||
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Annotation | LRRC8A(T48D):C conformation 1 LRR focus 1 unsharpened | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: LRRC8A(T48D):C conformation 1 LRR focus 1
File | emd_28903_half_map_1.map | ||||||||||||
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Annotation | LRRC8A(T48D):C conformation 1 LRR focus 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: LRRC8A(T48D):C conformation 1 LRR focus 1
File | emd_28903_half_map_2.map | ||||||||||||
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Annotation | LRRC8A(T48D):C conformation 1 LRR focus 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : LRRC8A:C
Entire | Name: LRRC8A:C |
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Components |
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-Supramolecule #1: LRRC8A:C
Supramolecule | Name: LRRC8A:C / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 620 KDa |
-Macromolecule #1: Volume-regulated anion channel subunit LRRC8A,Soluble cytochrome b562
Macromolecule | Name: Volume-regulated anion channel subunit LRRC8A,Soluble cytochrome b562 type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 105.544086 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MIPVTELRYF ADTQPAYRIL KPWWDVFTDY ISIVMLMIAV FGGTLQVDQD KMICLPCKWV TKDSCNDSFR GWAASSADLE DNWETLNDN LKVIEKADNA AQVKDALTKM RAAALDAQKA TPPKLEDKSP DSPEMKDFRH GFDILVGQID DALKLANEGK V KEAQAAAE ...String: MIPVTELRYF ADTQPAYRIL KPWWDVFTDY ISIVMLMIAV FGGTLQVDQD KMICLPCKWV TKDSCNDSFR GWAASSADLE DNWETLNDN LKVIEKADNA AQVKDALTKM RAAALDAQKA TPPKLEDKSP DSPEMKDFRH GFDILVGQID DALKLANEGK V KEAQAAAE QLKTTRNAYI QKYLDTGPTG IKYDLDRHQY NYVDAVCYEN RLHWFAKYFP YLVLLHTLIF LACSNFWFKF PR TSSKLEH FVSILLKCFD SPWTTRALSE TVVEESDPKP AFSKMNGSMD KKSSTVSEDV EATVPMLQRT KSRIEQGIVD RSE TGVLDK KEGEQAKALF EKVKKFRTHV EEGDIVYRLY MRQTIIKVIK FALIICYTVY YVHNIKFDVD CTVDIESLTG YRTY RCAHP LATLFKILAS FYISLVIFYG LICMYTLWWM LRRSLKKYSF ESIREESSYS DIPDVKNDFA FMLHLIDQYD PLYSK RFAV FLSEVSENKL RQLNLNNEWT LDKLRQRLTK NAQDKLELHL FMLSGIPDTV FDLVELEVLK LELIPDVTIP PSIAQL TGL KELWLYHTAA KIEAPALAFL RENLRALHIK FTDIKEIPLW IYSLKTLEEL HLTGNLSAEN NRYIVIDGLR ELKRLKV LR LKSNLSKLPQ VVTDVGVHLQ KLSINNEGTK LIVLNSLKKM VNLTELELIR CDLERIPHSI FSLHNLQEID LKDNNLKT I EEIISFQHLH RLTCLKLWYN HIAYIPIQIG NLTNLERLYL NRNKIEKIPT QLFYCRKLRY LDLSHNNLTF LPADIGLLQ NLQNLAVTAN RIEALPPELF QCRKLRALHL GNNVLQSLPS RVGELTNLTQ IELRGNRLEC LPVELGECPL LKRSGLVVEE DLFSTLPPE VKERLWRADK EQASNSLEVL FQ UniProtKB: Volume-regulated anion channel subunit LRRC8A, Soluble cytochrome b562, Volume-regulated anion channel subunit LRRC8A |
-Macromolecule #2: Volume-regulated anion channel subunit LRRC8C
Macromolecule | Name: Volume-regulated anion channel subunit LRRC8C / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 93.624594 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MIPVTEFRQF SEQQPAFRVL KPWWDVFTDY LSVAMLMIGV FGCTLQVMQD KIICLPKRVQ PAQNHSSVPN VSQAVISTTP LPPPKPSPT NPATVEMKGL KTDLDLQQYS FINQMCYERA LHWYAKYFPY LVLIHTLVFM LCSNFWFKFP GSSSKIEHFI S ILGKCFDS ...String: MIPVTEFRQF SEQQPAFRVL KPWWDVFTDY LSVAMLMIGV FGCTLQVMQD KIICLPKRVQ PAQNHSSVPN VSQAVISTTP LPPPKPSPT NPATVEMKGL KTDLDLQQYS FINQMCYERA LHWYAKYFPY LVLIHTLVFM LCSNFWFKFP GSSSKIEHFI S ILGKCFDS PWTTRALSEV SGEDSEEKDN RKNNMNRSGT IQSGPEGNLV RSQSLKSIPE KFVVDKSAAG ALDKKEGEQA KA LFEKVKK FRLHVEEGDI LYAMYVRQTV LKVIKFLIII AYNSALVSKV QFTVDCNVDI QDMTGYKNFS CNHTMAHLFS KLS FCYLCF VSIYGLTCLY TLYWLFYRSL REYSFEYVRQ ETGIDDIPDV KNDFAFMLHM IDQYDPLYSK RFAVFLSEVS ENKL KQLNL NNEWTPDKLR QKLQTNAHNR LELPLIMLSG LPDTVFEITE LQSLKLEIIK NVMIPATIAQ LDNLQELCLH QCSVK IHSA ALSFLKENLK VLSVKFDDMR ELPPWMYGLR NLEELYLVGS LSHDISKNVT LESLRDLKSL KILSIKSNVS KIPQAV VDV SSHLQKMCVH NDGTKLVMLN NLKKMTNLTE LELVHCDLER IPHAVFSLLS LQELDLKENN LKSIEEIVSF QHLRKLT VL KLWYNSIAYI PEHIKKLTSL ERLFFSHNKV EVLPSHLFLC NKIRYLDLSY NDIRFIPPEI GVLQSLQYFS ITCNKVES L PDELYFCKKL KTLKIGKNSL SVLSPKIGNL LFLSYLDIKG NHFEVLPPEL GDCRALKRAG LVVEDALFET LPSDVREQM KADSNSENLY FQG UniProtKB: Volume-regulated anion channel subunit LRRC8C |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 2 mg/mL |
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Buffer | pH: 7.4 |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.6 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 4.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 71566 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |