+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-30617 | |||||||||
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Title | human potassium-chloride co-transporter KCC4 | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information ammonium import across plasma membrane / potassium:chloride symporter activity / Cation-coupled Chloride cotransporters / chloride ion homeostasis / ammonium channel activity / potassium ion homeostasis / cell volume homeostasis / plasma membrane => GO:0005886 / potassium ion import across plasma membrane / monoatomic ion transport ...ammonium import across plasma membrane / potassium:chloride symporter activity / Cation-coupled Chloride cotransporters / chloride ion homeostasis / ammonium channel activity / potassium ion homeostasis / cell volume homeostasis / plasma membrane => GO:0005886 / potassium ion import across plasma membrane / monoatomic ion transport / chloride transmembrane transport / chemical synaptic transmission / synapse / protein kinase binding / protein-containing complex / plasma membrane Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
Authors | Xie Y / Chang S / Zhao C / Ye S / Guo J | |||||||||
Funding support | China, 1 items
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Citation | Journal: Sci Adv / Year: 2020 Title: Structures and an activation mechanism of human potassium-chloride cotransporters. Authors: Yuan Xie / Shenghai Chang / Cheng Zhao / Feng Wang / Si Liu / Jin Wang / Eric Delpire / Sheng Ye / Jiangtao Guo / Abstract: Potassium-chloride cotransporters KCC1 to KCC4 mediate the coupled export of potassium and chloride across the plasma membrane and play important roles in cell volume regulation, auditory system ...Potassium-chloride cotransporters KCC1 to KCC4 mediate the coupled export of potassium and chloride across the plasma membrane and play important roles in cell volume regulation, auditory system function, and γ-aminobutyric acid (GABA) and glycine-mediated inhibitory neurotransmission. Here, we present 2.9- to 3.6-Å resolution structures of full-length human KCC2, KCC3, and KCC4. All three KCCs adopt a similar overall architecture, a domain-swap dimeric assembly, and an inward-facing conformation. The structural and functional studies reveal that one unexpected N-terminal peptide binds at the cytosolic facing cavity and locks KCC2 and KCC4 at an autoinhibition state. The C-terminal domain (CTD) directly interacts with the N-terminal inhibitory peptide, and the relative motions between the CTD and the transmembrane domain (TMD) suggest that CTD regulates KCCs' activities by adjusting the autoinhibitory effect. These structures provide the first glimpse of full-length structures of KCCs and an autoinhibition mechanism among the amino acid-polyamine-organocation transporter superfamily. | |||||||||
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_30617.map.gz | 35 MB | EMDB map data format | |
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Header (meta data) | emd-30617-v30.xml emd-30617.xml | 10.7 KB 10.7 KB | Display Display | EMDB header |
Images | emd_30617.png | 52.3 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-30617 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-30617 | HTTPS FTP |
-Validation report
Summary document | emd_30617_validation.pdf.gz | 473.2 KB | Display | EMDB validaton report |
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Full document | emd_30617_full_validation.pdf.gz | 472.8 KB | Display | |
Data in XML | emd_30617_validation.xml.gz | 6 KB | Display | |
Data in CIF | emd_30617_validation.cif.gz | 6.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30617 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30617 | HTTPS FTP |
-Related structure data
Related structure data | 7d99MC 7d8zC 7d90C 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_30617.map.gz / Format: CCP4 / Size: 52.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 1.014 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : potassium-chloride cotransporter 4
Entire | Name: potassium-chloride cotransporter 4 |
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Components |
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-Supramolecule #1: potassium-chloride cotransporter 4
Supramolecule | Name: potassium-chloride cotransporter 4 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant cell: HEK293T |
-Macromolecule #1: potassium-chloride co-transporter KCC4
Macromolecule | Name: potassium-chloride co-transporter KCC4 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 123.069492 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: MDYKDDDDKH HHHHHENLYF QGMPTNFTVV PVEAHADGGG DETAERTEAP GTPEGPEPER PSPGDGNPRE NSPFLNNVEV EQESFFEGK NMALFEEEMD SNPMVSSLLN KLANYTNLSQ GVVEHEEDEE SRRREAKAPR MGTFIGVYLP CLQNILGVIL F LRLTWIVG ...String: MDYKDDDDKH HHHHHENLYF QGMPTNFTVV PVEAHADGGG DETAERTEAP GTPEGPEPER PSPGDGNPRE NSPFLNNVEV EQESFFEGK NMALFEEEMD SNPMVSSLLN KLANYTNLSQ GVVEHEEDEE SRRREAKAPR MGTFIGVYLP CLQNILGVIL F LRLTWIVG VAGVLESFLI VAMCCTCTML TAISMSAIAT NGVVPAGGSY YMISRSLGPE FGGAVGLCFY LGTTFAGAMY IL GTIEIFL TYISPGAAIF QAEAAGGEAA AMLHNMRVYG TCTLVLMALV VFVGVKYVNK LALVFLACVV LSILAIYAGV IKS AFDPPD IPVCLLGNRT LSRRSFDACV KAYGIHNNSA TSALWGLFCN GSQPSAACDE YFIQNNVTEI QGIPGAASGV FLEN LWSTY AHAGAFVEKK GVPSVPVAEE SRASALPYVL TDIAASFTLL VGIYFPSVTG IMAGSNRSGD LKDAQKSIPT GTILA IVTT SFIYLSCIVL FGACIEGVVL RDKFGEALQG NLVIGMLAWP SPWVIVIGSF FSTCGAGLQS LTGAPRLLQA IARDGI VPF LQVFGHGKAN GEPTWALLLT VLICETGILI ASLDSVAPIL SMFFLMCYLF VNLACAVQTL LRTPNWRPRF KFYHWTL SF LGMSLCLALM FICSWYYALS AMLIAGCIYK YIEYRGAEKE WGDGIRGLSL NAARYALLRV EHGPPHTKNW RPQVLVML N LDAEQAVKHP RLLSFTSQLK AGKGLTIVGS VLEGTYLDKH MEAQRAEENI RSLMSTEKTK GFCQLVVSSS LRDGMSHLI QSAGLGGLKH NTVLMAWPAS WKQEDNPFSW KNFVDTVRDT TAAHQALLVA KNVDSFPQNQ ERFGGGHIDV WWIVHDGGML MLLPFLLRQ HKVWRKCRMR IFTVAQVDDN SIQMKKDLQM FLYHLRISAE VEVVEMVEND ISAFTYERTL MMEQRSQMLK Q MQLSKNEQ EREAQLIHDR NTASHTAAAA RTQAPPTPDK VQMTWTREKL IAEKYRSRDT SLSGFKDLFS MKPDQSNVRR MH TAVKLNG VVLNKSQDAQ LVLLNMPGPP KNRQGDENYM EFLEVLTEGL NRVLLVRGGG REVITIYSWS HPQFEK |
-Macromolecule #3: POTASSIUM ION
Macromolecule | Name: POTASSIUM ION / type: ligand / ID: 3 / Number of copies: 2 / Formula: K |
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Molecular weight | Theoretical: 39.098 Da |
-Macromolecule #4: CHLORIDE ION
Macromolecule | Name: CHLORIDE ION / type: ligand / ID: 4 / Number of copies: 4 / Formula: CL |
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Molecular weight | Theoretical: 35.453 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 62.0 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 |
-Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 95572 |
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Initial angle assignment | Type: ANGULAR RECONSTITUTION |
Final angle assignment | Type: ANGULAR RECONSTITUTION |