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- EMDB-11802: Structure of Human Potassium Chloride Transporter KCC1 in NaCl (S... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-11802 | |||||||||
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Title | Structure of Human Potassium Chloride Transporter KCC1 in NaCl (Subclass 1) | |||||||||
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Function / homology | ![]() 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 / potassium ion import across plasma membrane / monoatomic ion transport / chloride transmembrane 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 / potassium ion import across plasma membrane / monoatomic ion transport / chloride transmembrane transport / potassium ion transmembrane transport / chemical synaptic transmission / lysosomal membrane / synapse / protein kinase binding / ATP binding / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.72 Å | |||||||||
![]() | Ebenhoch R / Chi G / Man H / Wang D / McKinley G / Mukhopadhyay SMM / MacLean EM / Chalk R / Moreau C / Snee M ...Ebenhoch R / Chi G / Man H / Wang D / McKinley G / Mukhopadhyay SMM / MacLean EM / Chalk R / Moreau C / Snee M / Bohstedt T / Singh NK / Abrusci P / Liko I / Tehan BG / Almeida FG / Arrowsmith CH / Tang H / Robinson CV / Bountra C / Edwards AM / Marsden BD / Burgess-Brown NA / Duerr KL / Structural Genomics Consortium (SGC) | |||||||||
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![]() | ![]() Title: Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters. Authors: Gamma Chi / Rebecca Ebenhoch / Henry Man / Haiping Tang / Laurence E Tremblay / Gabriella Reggiano / Xingyu Qiu / Tina Bohstedt / Idlir Liko / Fernando G Almeida / Alexandre P Garneau / Dong ...Authors: Gamma Chi / Rebecca Ebenhoch / Henry Man / Haiping Tang / Laurence E Tremblay / Gabriella Reggiano / Xingyu Qiu / Tina Bohstedt / Idlir Liko / Fernando G Almeida / Alexandre P Garneau / Dong Wang / Gavin McKinley / Christophe P Moreau / Kiran D Bountra / Patrizia Abrusci / Shubhashish M M Mukhopadhyay / Alejandra Fernandez-Cid / Samira Slimani / Julie L Lavoie / Nicola A Burgess-Brown / Ben Tehan / Frank DiMaio / Ali Jazayeri / Paul Isenring / Carol V Robinson / Katharina L Dürr / ![]() ![]() ![]() Abstract: Potassium-coupled chloride transporters (KCCs) play crucial roles in regulating cell volume and intracellular chloride concentration. They are characteristically inhibited under isotonic conditions ...Potassium-coupled chloride transporters (KCCs) play crucial roles in regulating cell volume and intracellular chloride concentration. They are characteristically inhibited under isotonic conditions via phospho-regulatory sites located within the cytoplasmic termini. Decreased inhibitory phosphorylation in response to hypotonic cell swelling stimulates transport activity, and dysfunction of this regulatory process has been associated with various human diseases. Here, we present cryo-EM structures of human KCC3b and KCC1, revealing structural determinants for phospho-regulation in both N- and C-termini. We show that phospho-mimetic KCC3b is arrested in an inward-facing state in which intracellular ion access is blocked by extensive contacts with the N-terminus. In another mutant with increased isotonic transport activity, KCC1Δ19, this interdomain interaction is absent, likely due to a unique phospho-regulatory site in the KCC1 N-terminus. Furthermore, we map additional phosphorylation sites as well as a previously unknown ATP/ADP-binding pocket in the large C-terminal domain and show enhanced thermal stabilization of other CCCs by adenine nucleotides. These findings provide fundamentally new insights into the complex regulation of KCCs and may unlock innovative strategies for drug development. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 97.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18.3 KB 18.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.7 KB | Display | ![]() |
Images | ![]() | 163.6 KB | ||
Masks | ![]() | 103 MB | ![]() | |
Others | ![]() ![]() | 95.7 MB 95.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7aiqMC ![]() 6y5vC ![]() 7ainC ![]() 7aioC ![]() 7aipC ![]() 7airC ![]() 7ngbC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 0.85 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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-Half map: #2
File | emd_11802_half_map_1.map | ||||||||||||
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-Half map: #1
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Density Histograms |
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Sample components
-Entire : Homodimeric complex of human potassium chloride transporter KCC1 ...
Entire | Name: Homodimeric complex of human potassium chloride transporter KCC1 in complex with ATP and magnesium |
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Components |
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-Supramolecule #1: Homodimeric complex of human potassium chloride transporter KCC1 ...
Supramolecule | Name: Homodimeric complex of human potassium chloride transporter KCC1 in complex with ATP and magnesium type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Molecular weight | Theoretical: 240 KDa |
-Macromolecule #1: Solute carrier family 12 member 4
Macromolecule | Name: Solute carrier family 12 member 4 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 119.610281 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MLEGLSWVDY GERAELDDSD GHGNHRESSP FLSPLEASRG IDYYDRNLAL FEEELDIRPK VSSLLGKLVS YTNLTQGAKE HEEAESGEG TRRRAAEAPS MGTLMGVYLP CLQNIFGVIL FLRLTWMVGT AGVLQALLIV LICCCCTLLT AISMSAIATN G VVPAGGSY ...String: MLEGLSWVDY GERAELDDSD GHGNHRESSP FLSPLEASRG IDYYDRNLAL FEEELDIRPK VSSLLGKLVS YTNLTQGAKE HEEAESGEG TRRRAAEAPS MGTLMGVYLP CLQNIFGVIL FLRLTWMVGT AGVLQALLIV LICCCCTLLT AISMSAIATN G VVPAGGSY FMISRSLGPE FGGAVGLCFY LGTTFAAAMY ILGAIEILLT YIAPPAAIFY PSGAHDTSNA TLNNMRVYGT IF LTFMTLV VFVGVKYVNK FASLFLACVI ISILSIYAGG IKSIFDPPVF PVCMLGNRTL SRDQFDICAK TAVVDNETVA TQL WSFFCH SPNLTTDSCD PYFMLNNVTE IPGIPGAAAG VLQENLWSAY LEKGDIVEKH GLPSADAPSL KESLPLYVVA DIAT SFTVL VGIFFPSVTG IMAGSNRSGD LRDAQKSIPV GTILAIITTS LVYFSSVVLF GACIEGVVLR DKYGDGVSRN LVVGT LAWP SPWVIVIGSF FSTCGAGLQS LTGAPRLLQA IAKDNIIPFL RVFGHGKVNG EPTWALLLTA LIAELGILIA SLDMVA PIL SMFFLMCYLF VNLACAVQTL LRTPNWRPRF KYYHWALSFL GMSLCLALMF VSSWYYALVA MLIAGMIYKY IEYQGAE KE WGDGIRGLSL SAARYALLRL EEGPPHTKNW RPQLLVLLKL DEDLHVKYPR LLTFASQLKA GKGLTIVGSV IQGSFLES Y GEAQAAEQTI KNMMEIEKVK GFCQVVVASK VREGLAHLIQ SCGLGGMRHN SVVLGWPYGW RQSEDPRAWK TFIDTVRCT TAAHLALLVP KNIAFYPSNH ERYLEGHIDV WWIVHDGGML MLLPFLLRQH KVWRKCRMRI FTVAQMDDNS IQMKKDLAVF LYHLRLEAE VEVVEMHNSD ISAYTYERTL MMEQRSQMLR QMRLTKTERE REAQLVKDRH SALRLESLYS DEEDESAVGA D KIQMTWTR DKYMTETWDP SHAPDNFREL VHIKPDQSNV RRMHTAVKLN EVIVTRSHDA RLVLLNMPGP PRNSEGDENY ME FLEVLTE GLERVLLVRG GGREVITIYS AENLYFQ |
-Macromolecule #3: 2-acetamido-2-deoxy-beta-D-glucopyranose
Macromolecule | Name: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 3 / Number of copies: 2 / Formula: NAG |
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Molecular weight | Theoretical: 221.208 Da |
Chemical component information | ![]() ChemComp-NAG: |
-Macromolecule #4: ADENOSINE-5'-TRIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 4 / Number of copies: 2 / Formula: ATP |
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Molecular weight | Theoretical: 507.181 Da |
Chemical component information | ![]() ChemComp-ATP: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Details | Model from reference map/model fitted with Chimera, refined with Coot and then with Phenix. |
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Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
Output model | ![]() PDB-7aiq: |