+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-0471 | |||||||||
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Title | Structure of the NKCC1 CTD | |||||||||
Map data | Cytosolic domain | |||||||||
Sample |
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Keywords | membrane protein | |||||||||
Function / homology | Function and homology information swim bladder inflation / Cation-coupled Chloride cotransporters / chloride:monoatomic cation symporter activity / sodium:potassium:chloride symporter activity / sodium ion homeostasis / ear development / ammonium transmembrane transport / ammonium channel activity / chloride ion homeostasis / potassium ion homeostasis ...swim bladder inflation / Cation-coupled Chloride cotransporters / chloride:monoatomic cation symporter activity / sodium:potassium:chloride symporter activity / sodium ion homeostasis / ear development / ammonium transmembrane transport / ammonium channel activity / chloride ion homeostasis / potassium ion homeostasis / cell volume homeostasis / inner ear morphogenesis / potassium ion import across plasma membrane / sodium ion transmembrane transport / chloride transmembrane transport / basolateral plasma membrane / apical plasma membrane / identical protein binding Similarity search - Function | |||||||||
Biological species | Danio rerio (zebrafish) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | Feng L / Liao MF | |||||||||
Citation | Journal: Nature / Year: 2019 Title: Structure and mechanism of the cation-chloride cotransporter NKCC1. Authors: Thomas A Chew / Benjamin J Orlando / Jinru Zhang / Naomi R Latorraca / Amy Wang / Scott A Hollingsworth / Dong-Hua Chen / Ron O Dror / Maofu Liao / Liang Feng / Abstract: Cation-chloride cotransporters (CCCs) mediate the electroneutral transport of chloride, potassium and/or sodium across the membrane. They have critical roles in regulating cell volume, controlling ...Cation-chloride cotransporters (CCCs) mediate the electroneutral transport of chloride, potassium and/or sodium across the membrane. They have critical roles in regulating cell volume, controlling ion absorption and secretion across epithelia, and maintaining intracellular chloride homeostasis. These transporters are primary targets for some of the most commonly prescribed drugs. Here we determined the cryo-electron microscopy structure of the Na-K-Cl cotransporter NKCC1, an extensively studied member of the CCC family, from Danio rerio. The structure defines the architecture of this protein family and reveals how cytosolic and transmembrane domains are strategically positioned for communication. Structural analyses, functional characterizations and computational studies reveal the ion-translocation pathway, ion-binding sites and key residues for transport activity. These results provide insights into ion selectivity, coupling and translocation, and establish a framework for understanding the physiological functions of CCCs and interpreting disease-related mutations. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_0471.map.gz | 37.4 MB | EMDB map data format | |
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Header (meta data) | emd-0471-v30.xml emd-0471.xml | 12.4 KB 12.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_0471_fsc.xml | 9.2 KB | Display | FSC data file |
Images | emd_0471.png | 82.6 KB | ||
Filedesc metadata | emd-0471.cif.gz | 5.6 KB | ||
Others | emd_0471_additional.map.gz | 25.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-0471 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-0471 | HTTPS FTP |
-Validation report
Summary document | emd_0471_validation.pdf.gz | 605.4 KB | Display | EMDB validaton report |
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Full document | emd_0471_full_validation.pdf.gz | 604.9 KB | Display | |
Data in XML | emd_0471_validation.xml.gz | 9.9 KB | Display | |
Data in CIF | emd_0471_validation.cif.gz | 12.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0471 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0471 | HTTPS FTP |
-Related structure data
Related structure data | 6npjMC 0470C 0472C 0473C 0474C 0475C 6nphC 6npkC 6nplC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_0471.map.gz / Format: CCP4 / Size: 40.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cytosolic domain | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.055 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Cytosolic domain
File | emd_0471_additional.map | ||||||||||||
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Annotation | Cytosolic domain | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : cytosolic domain
Entire | Name: cytosolic domain |
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Components |
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-Supramolecule #1: cytosolic domain
Supramolecule | Name: cytosolic domain / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Danio rerio (zebrafish) |
Molecular weight | Theoretical: 150 KDa |
-Macromolecule #1: Sodium-potassium-chloride cotransporter 1
Macromolecule | Name: Sodium-potassium-chloride cotransporter 1 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Danio rerio (zebrafish) |
Molecular weight | Theoretical: 49.444598 KDa |
Recombinant expression | Organism: Autographa californica nucleopolyhedrovirus |
Sequence | String: WGSSTQALTY HQALTHSLQL CGVADHIKTF RPQCLVMTGA PNSRPAILHL VHAFTKNVGL MLCGHVRISS RRPNFKELNS DMLRYQRWL LNNNSKAFYT CVVAEDLRQG TQYMLQAAGL GRLRPNTLVI GFKNDWRIGD IKEVETYINL IHDAFDFQYG V VILRLREG ...String: WGSSTQALTY HQALTHSLQL CGVADHIKTF RPQCLVMTGA PNSRPAILHL VHAFTKNVGL MLCGHVRISS RRPNFKELNS DMLRYQRWL LNNNSKAFYT CVVAEDLRQG TQYMLQAAGL GRLRPNTLVI GFKNDWRIGD IKEVETYINL IHDAFDFQYG V VILRLREG LDISHIQGQD DSSGMKDVVV SVDISKDSDG DSSKPSSKAT SVQNSPAVQK DKKSPTVPLN VADQRLLDSQ QF QQKQGKG TVDVWWLFDD GGLTLLIPYL IANKKKWKDC KIRVFIGGKI NRIDHDRRAM ATLLSKFRID FSDITVLGDI NTK PKSEGL TEFAEMIEPY KLREDDMEQE AAEKLKSEEP WRITANELEL YKAKGNRQIR LNELLKEHSS TANLIVMSMP LARK GAVSS ALYMAWLDTL SKDLPPILLV RGNHQSVLTF YS UniProtKB: Solute carrier family 12 member 2 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 7.9 mg/mL |
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Buffer | pH: 8 |
Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 53.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |