[English] 日本語
Yorodumi- EMDB-13212: Cryo-EM structure of homomeric LRRC8A Volume-Regulated Anion Chan... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13212 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of homomeric LRRC8A Volume-Regulated Anion Channel in complex with synthetic nanobody Sb4 at 1:1 ratio | |||||||||
Map data | ||||||||||
Sample |
| |||||||||
Function / homology | Function and homology information 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 / cell volume homeostasis / protein hexamerization ...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 / cell volume homeostasis / protein hexamerization / monoatomic anion transport / response to osmotic stress / monoatomic ion channel complex / intracellular glucose homeostasis / positive regulation of myoblast differentiation / chloride transmembrane transport / positive regulation of insulin secretion / spermatogenesis / lysosomal membrane / cell surface / identical protein binding / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | Mus musculus (house mouse) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.67 Å | |||||||||
Authors | Deneka D / Rutz S / Sawicka M | |||||||||
Funding support | 1 items
| |||||||||
Citation | Journal: Nat Commun / Year: 2021 Title: Allosteric modulation of LRRC8 channels by targeting their cytoplasmic domains. Authors: Dawid Deneka / Sonja Rutz / Cedric A J Hutter / Markus A Seeger / Marta Sawicka / Raimund Dutzler / Abstract: Members of the LRRC8 family form heteromeric assemblies, which function as volume-regulated anion channels. These modular proteins consist of a transmembrane pore and cytoplasmic leucine-rich repeat ...Members of the LRRC8 family form heteromeric assemblies, which function as volume-regulated anion channels. These modular proteins consist of a transmembrane pore and cytoplasmic leucine-rich repeat (LRR) domains. Despite their known molecular architecture, the mechanism of activation and the role of the LRR domains in this process has remained elusive. Here we address this question by generating synthetic nanobodies, termed sybodies, which target the LRR domain of the obligatory subunit LRRC8A. We use these binders to investigate their interaction with homomeric LRRC8A channels by cryo-electron microscopy and the consequent effect on channel activation by electrophysiology. The five identified sybodies either inhibit or enhance activity by binding to distinct epitopes of the LRR domain, thereby altering channel conformations. In combination, our work provides a set of specific modulators of LRRC8 proteins and reveals the role of their cytoplasmic domains as regulators of channel activity by allosteric mechanisms. | |||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_13212.map.gz | 11.2 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-13212-v30.xml emd-13212.xml | 8.4 KB 8.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_13212_fsc.xml | 12 KB | Display | FSC data file |
Images | emd_13212.png | 27.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13212 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13212 | HTTPS FTP |
-Validation report
Summary document | emd_13212_validation.pdf.gz | 345.1 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_13212_full_validation.pdf.gz | 344.6 KB | Display | |
Data in XML | emd_13212_validation.xml.gz | 12.5 KB | Display | |
Data in CIF | emd_13212_validation.cif.gz | 16.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13212 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13212 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_13212.map.gz / Format: CCP4 / Size: 144.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Voxel size | X=Y=Z: 1.302 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : Homomeric LRRC8A Volume-Regulated Anion Channel in complex with s...
Entire | Name: Homomeric LRRC8A Volume-Regulated Anion Channel in complex with synthetic nanobody Sb4 at 1:1 ratio |
---|---|
Components |
|
-Supramolecule #1: Homomeric LRRC8A Volume-Regulated Anion Channel in complex with s...
Supramolecule | Name: Homomeric LRRC8A Volume-Regulated Anion Channel in complex with synthetic nanobody Sb4 at 1:1 ratio type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
---|---|
Source (natural) | Organism: Mus musculus (house mouse) |
Recombinant expression | Organism: Homo sapiens (human) |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
---|---|
Vitrification | Cryogen name: ETHANE-PROPANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 67.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |