+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-30543 | |||||||||
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Title | Mouse KCC2 | |||||||||
Map data | ||||||||||
Sample |
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Keywords | transporter / MEMBRANE PROTEIN | |||||||||
Function / homology | Function and homology information Cation-coupled Chloride cotransporters / ammonium transmembrane transport => GO:0072488 / thermosensory behavior / intracellular pH reduction / potassium:chloride symporter activity / hypotonic response / intracellular chloride ion homeostasis / response to xenobiotic stimulus => GO:0009410 / chloride ion homeostasis / intracellular monoatomic ion homeostasis ...Cation-coupled Chloride cotransporters / ammonium transmembrane transport => GO:0072488 / thermosensory behavior / intracellular pH reduction / potassium:chloride symporter activity / hypotonic response / intracellular chloride ion homeostasis / response to xenobiotic stimulus => GO:0009410 / chloride ion homeostasis / intracellular monoatomic ion homeostasis / dendritic spine development / ammonium channel activity / potassium ion homeostasis / cell volume homeostasis / chloride transport / chloride transmembrane transporter activity / plasma membrane => GO:0005886 / potassium ion import across plasma membrane / monoatomic ion transport / chloride transmembrane transport / dendrite membrane / potassium ion transport / learning / dendritic shaft / cell periphery / multicellular organism growth / chemical synaptic transmission / perikaryon / membrane => GO:0016020 / neuron projection / neuronal cell body / synapse / protein kinase binding / plasma membrane Similarity search - Function | |||||||||
Biological species | Mus musculus (house mouse) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | Zhang S / Yang M | |||||||||
Citation | Journal: Commun Biol / Year: 2021 Title: The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2. Authors: Sensen Zhang / Jun Zhou / Yuebin Zhang / Tianya Liu / Perrine Friedel / Wei Zhuo / Suma Somasekharan / Kasturi Roy / Laixing Zhang / Yang Liu / Xianbin Meng / Haiteng Deng / Wenwen Zeng / ...Authors: Sensen Zhang / Jun Zhou / Yuebin Zhang / Tianya Liu / Perrine Friedel / Wei Zhuo / Suma Somasekharan / Kasturi Roy / Laixing Zhang / Yang Liu / Xianbin Meng / Haiteng Deng / Wenwen Zeng / Guohui Li / Biff Forbush / Maojun Yang / Abstract: NKCC and KCC transporters mediate coupled transport of Na+K+Cl and K+Cl across the plasma membrane, thus regulating cell Cl concentration and cell volume and playing critical roles in transepithelial ...NKCC and KCC transporters mediate coupled transport of Na+K+Cl and K+Cl across the plasma membrane, thus regulating cell Cl concentration and cell volume and playing critical roles in transepithelial salt and water transport and in neuronal excitability. The function of these transporters has been intensively studied, but a mechanistic understanding has awaited structural studies of the transporters. Here, we present the cryo-electron microscopy (cryo-EM) structures of the two neuronal cation-chloride cotransporters human NKCC1 (SLC12A2) and mouse KCC2 (SLC12A5), along with computational analysis and functional characterization. These structures highlight essential residues in ion transport and allow us to propose mechanisms by which phosphorylation regulates transport activity. | |||||||||
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_30543.map.gz | 9.3 MB | EMDB map data format | |
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Header (meta data) | emd-30543-v30.xml emd-30543.xml | 10.1 KB 10.1 KB | Display Display | EMDB header |
Images | emd_30543.png | 68.8 KB | ||
Filedesc metadata | emd-30543.cif.gz | 5.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-30543 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-30543 | HTTPS FTP |
-Validation report
Summary document | emd_30543_validation.pdf.gz | 368.8 KB | Display | EMDB validaton report |
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Full document | emd_30543_full_validation.pdf.gz | 368.4 KB | Display | |
Data in XML | emd_30543_validation.xml.gz | 6.5 KB | Display | |
Data in CIF | emd_30543_validation.cif.gz | 7.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30543 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30543 | HTTPS FTP |
-Related structure data
Related structure data | 7d14MC 7d10C 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_30543.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 1.091 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 : mouse KCC2
Entire | Name: mouse KCC2 |
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Components |
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-Supramolecule #1: mouse KCC2
Supramolecule | Name: mouse KCC2 / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Mus musculus (house mouse) |
-Macromolecule #1: Solute carrier family 12 member 5
Macromolecule | Name: Solute carrier family 12 member 5 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Mus musculus (house mouse) |
Molecular weight | Theoretical: 126.402328 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: MSRRFTVTSL PPAASAASAD PESRRHSVAD PRRLPREDVK GDGNPKESSP FINSTDTEKG REYDGRNMAL FEEEMDTSPM VSSLLSGLA NYTNLPQGSR EHEEAENNEG GKKKPVQAPR MGTFMGVYLP CLQNIFGVIL FLRLTWVVGI AGIMESFCMV F ICCSCTML ...String: MSRRFTVTSL PPAASAASAD PESRRHSVAD PRRLPREDVK GDGNPKESSP FINSTDTEKG REYDGRNMAL FEEEMDTSPM VSSLLSGLA NYTNLPQGSR EHEEAENNEG GKKKPVQAPR MGTFMGVYLP CLQNIFGVIL FLRLTWVVGI AGIMESFCMV F ICCSCTML TAISMSAIAT NGVVPAGGSY YMISRSLGPE FGGAVGLCFY LGTTFAGAMY ILGTIEILLA YLFPAMAIFK AE DASGEAA AMLNNMRVYG TCVLTCMATV VFVGVKYVNK FALVFLGCVI LSILAIYAGV IKSAFDPPNF PICLLGNRTL SRH GFDVCA KLAWEGNETV TTRLWGLFCS SRLLNATCDE YFTRNNVTEI QGIPGAASGL IKENLWSSYL TKGVIVERRG MPSV GLADG TPVDMDHPYV FSDMTSYFTL LVGIYFPSVT GIMAGSNRSG DLRDAQKSIP TGTILAIATT SAVYISSVVL FGACI EGVV LRDKFGEAVN GNLVVGTLAW PSPWVIVIGS FFSTCGAGLQ SLTGAPRLLQ AISRDGIVPF LQVFGHGKAN GEPTWA LLL TACICEIGIL IASLDEVAPI LSMFFLMCYM FVNLACAVQT LLRTPNWRPR FRYYHWTLSF LGMSLCLALM FICSWYY AL VAMLIAGLIY KYIEYRGAEK EWGDGIRGLS LSAARYALLR LEEGPPHTKN WRPQLLVLVR VDQDQNVVHP QLLSLTSQ L KAGKGLTIVG SVLEGTFLDN HPQAQRAEES IRRLMEAEKV KGFCQVVISS NLRDGVSHLI QSGGLGGLQH NTVLVGWPR NWRQKEDHQT WRNFIELVRE TTAGHLALLV TKNVSMFPGN PERFSEGSID VWWIVHDGGM LMLLPFLLRH HKVWRKCKMR IFTVAQMDD NSIQMKKDLT TFLYHLRITA EVEVVEMHES DISAYTYEKT LVMEQRSQIL KQMHLTKNER EREIQSITDE S RGSIRRKN PANPRLRLNV PEETACDNEE KPEEEVQLIH DQSAPSCPSS SPSPGEEPEG ERETDPEVHL TWTKDKSVAE KN KGPSPVS SEGIKDFFSM KPEWENLNQS NVRRMHTAVR LNEVIVNKSR DAKLVLLNMP GPPRNRNGDE NYMEFLEVLT EQL DRVMLV RGGGREVITI YS UniProtKB: Solute carrier family 12 member 5 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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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: 50.0 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
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 66000 |
Initial angle assignment | Type: ANGULAR RECONSTITUTION |
Final angle assignment | Type: ANGULAR RECONSTITUTION |