+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21846 | |||||||||
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Title | TASK2 in MSP1D1 lipid nanodisc at pH 8.5 | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information Phase 4 - resting membrane potential / stabilization of membrane potential / regulation of resting membrane potential / potassium ion leak channel activity / potassium ion export across plasma membrane / outward rectifier potassium channel activity / potassium ion import across plasma membrane / voltage-gated potassium channel activity / potassium channel activity / potassium ion transmembrane transport / plasma membrane Similarity search - Function | |||||||||
Biological species | Mus musculus (house mouse) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.52 Å | |||||||||
Authors | Li B / Brohawn SG | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nature / Year: 2020 Title: Structural basis for pH gating of the two-pore domain K channel TASK2. Authors: Baobin Li / Robert A Rietmeijer / Stephen G Brohawn / Abstract: TASK2 (also known as KCNK5) channels generate pH-gated leak-type K currents to control cellular electrical excitability. TASK2 is involved in the regulation of breathing by chemosensory neurons of ...TASK2 (also known as KCNK5) channels generate pH-gated leak-type K currents to control cellular electrical excitability. TASK2 is involved in the regulation of breathing by chemosensory neurons of the retrotrapezoid nucleus in the brainstem and pH homeostasis by kidney proximal tubule cells. These roles depend on channel activation by intracellular and extracellular alkalization, but the mechanistic basis for TASK2 gating by pH is unknown. Here we present cryo-electron microscopy structures of Mus musculus TASK2 in lipid nanodiscs in open and closed conformations. We identify two gates, distinct from previously observed K channel gates, controlled by stimuli on either side of the membrane. Intracellular gating involves lysine protonation on inner helices and the formation of a protein seal between the cytoplasm and the channel. Extracellular gating involves arginine protonation on the channel surface and correlated conformational changes that displace the K-selectivity filter to render it nonconductive. These results explain how internal and external protons control intracellular and selectivity filter gates to modulate TASK2 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_21846.map.gz | 28.4 MB | EMDB map data format | |
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Header (meta data) | emd-21846-v30.xml emd-21846.xml | 14.9 KB 14.9 KB | Display Display | EMDB header |
Images | emd_21846.png | 78.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21846 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21846 | HTTPS FTP |
-Validation report
Summary document | emd_21846_validation.pdf.gz | 188.4 KB | Display | EMDB validaton report |
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Full document | emd_21846_full_validation.pdf.gz | 188 KB | Display | |
Data in XML | emd_21846_validation.xml.gz | 502 B | Display | |
Data in CIF | emd_21846_validation.cif.gz | 374 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21846 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21846 | HTTPS FTP |
-Related structure data
Related structure data | 6wm0MC 6wlvC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | |
EM raw data | EMPIAR-10422 (Title: TASK2 in MSP1D1 lipid nanodisc at pH8.5 / Data size: 2.3 TB Data #1: multi-frame micrographs for ion channel TASK2 in open state [micrographs - multiframe]) |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21846.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.137 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 : TASK2 in MSP1D1 lipid nanodisc at pH 8.5
Entire | Name: TASK2 in MSP1D1 lipid nanodisc at pH 8.5 |
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Components |
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-Supramolecule #1: TASK2 in MSP1D1 lipid nanodisc at pH 8.5
Supramolecule | Name: TASK2 in MSP1D1 lipid nanodisc at pH 8.5 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Mus musculus (house mouse) |
Recombinant expression | Organism: Komagataella pastoris (fungus) |
Molecular weight | Theoretical: 77 KDa |
-Macromolecule #1: Potassium channel TASK2
Macromolecule | Name: Potassium channel TASK2 / 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: 38.661832 KDa |
Recombinant expression | Organism: Komagataella pastoris (fungus) |
Sequence | String: MVDRGPLLTS AIIFYLAIGA AIFEVLEEPH WKEAKKNYYT QKLHLLKEFP CLSQEGLDKI LQVVSDAADQ GVAITGNQTF NNWNWPNAM IFAATVITTI GYGNVAPKTP AGRLFCVFYG LFGVPLCLTW ISALGKFFGG RAKRLGQFLT RRGVSLRKAQ I TCTAIFIV ...String: MVDRGPLLTS AIIFYLAIGA AIFEVLEEPH WKEAKKNYYT QKLHLLKEFP CLSQEGLDKI LQVVSDAADQ GVAITGNQTF NNWNWPNAM IFAATVITTI GYGNVAPKTP AGRLFCVFYG LFGVPLCLTW ISALGKFFGG RAKRLGQFLT RRGVSLRKAQ I TCTAIFIV WGVLVHLVIP PFVFMVTEEW NYIEGLYYSF ITISTIGFGD FVAGVNPSAN YHALYRYFVE LWIYLGLAWL SL FVNWKVS MFVEVHKAIK KRRRRRKESF ESSPHSRKAL QMAGSTASKD VNIFSFLSKK EETYNDLIKQ IGKKAMKTSG GGE RVPGPG HGLGPQGDRS NSLEVLFQ |
-Macromolecule #2: POTASSIUM ION
Macromolecule | Name: POTASSIUM ION / type: ligand / ID: 2 / Number of copies: 5 / Formula: K |
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Molecular weight | Theoretical: 39.098 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 1.2 mg/mL | ||||||||||||
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Buffer | pH: 8.5 Component:
Details: pH at 277K | ||||||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 12.0 nm / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV Details: 3uL sample, incubate 5 seconds, blot 3 seconds, blot force 1. |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Digitization - Dimensions - Width: 11520 pixel / Digitization - Dimensions - Height: 8184 pixel / Number grids imaged: 1 / Number real images: 2814 / Average exposure time: 0.11 sec. / Average electron dose: 1.00925 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: OTHER / Overall B value: 151.61 |
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Output model | PDB-6wm0: |