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Yorodumi- EMDB-9024: The voltage-activated Kv1.2-2.1 paddle chimera channel in lipid n... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9024 | |||||||||
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Title | The voltage-activated Kv1.2-2.1 paddle chimera channel in lipid nanodiscs | |||||||||
Map data | Kv1.2-2.1 paddle chimera in lipid nanodiscs, entire protein | |||||||||
Sample |
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Keywords | MEMBRANE PROTEIN / transport protein / potassium channel / lipid nanodisc | |||||||||
Function / homology | Function and homology information optic nerve structural organization / pinceau fiber / regulation of action potential / methylglyoxal reductase (NADPH) (acetol producing) activity / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / NADPH oxidation / paranodal junction / regulation of circadian sleep/wake cycle, non-REM sleep ...optic nerve structural organization / pinceau fiber / regulation of action potential / methylglyoxal reductase (NADPH) (acetol producing) activity / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / NADPH oxidation / paranodal junction / regulation of circadian sleep/wake cycle, non-REM sleep / potassium ion export across plasma membrane / axon initial segment / regulation of protein localization to cell surface / corpus callosum development / voltage-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / aldo-keto reductase (NADPH) activity / delayed rectifier potassium channel activity / outward rectifier potassium channel activity / juxtaparanode region of axon / Oxidoreductases; Acting on the CH-OH group of donors; With NAD+ or NADP+ as acceptor / regulation of potassium ion transmembrane transport / myoblast differentiation / Neutrophil degranulation / neuromuscular process / optic nerve development / action potential / regulation of dopamine secretion / neuronal cell body membrane / kinesin binding / lamellipodium membrane / calyx of Held / voltage-gated potassium channel activity / neuronal action potential / potassium channel regulator activity / hematopoietic progenitor cell differentiation / voltage-gated potassium channel complex / axon terminus / potassium ion transmembrane transport / sensory perception of pain / protein localization to plasma membrane / postsynaptic density membrane / potassium ion transport / protein homooligomerization / cerebral cortex development / cytoplasmic side of plasma membrane / lamellipodium / presynaptic membrane / perikaryon / postsynaptic membrane / transmembrane transporter binding / postsynaptic density / cytoskeleton / endosome / neuron projection / protein heterodimerization activity / axon / neuronal cell body / glutamatergic synapse / dendrite / endoplasmic reticulum membrane / protein-containing complex binding / membrane / plasma membrane / cytosol Similarity search - Function | |||||||||
Biological species | Rattus norvegicus (Norway rat) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
Authors | Matthies D / Bae C | |||||||||
Citation | Journal: Elife / Year: 2018 Title: Single-particle cryo-EM structure of a voltage-activated potassium channel in lipid nanodiscs. Authors: Doreen Matthies / Chanhyung Bae / Gilman Es Toombes / Tara Fox / Alberto Bartesaghi / Sriram Subramaniam / Kenton Jon Swartz / Abstract: Voltage-activated potassium (Kv) channels open to conduct K ions in response to membrane depolarization, and subsequently enter non-conducting states through distinct mechanisms of inactivation. X- ...Voltage-activated potassium (Kv) channels open to conduct K ions in response to membrane depolarization, and subsequently enter non-conducting states through distinct mechanisms of inactivation. X-ray structures of detergent-solubilized Kv channels appear to have captured an open state even though a non-conducting C-type inactivated state would predominate in membranes in the absence of a transmembrane voltage. However, structures for a voltage-activated ion channel in a lipid bilayer environment have not yet been reported. Here we report the structure of the Kv1.2-2.1 paddle chimera channel reconstituted into lipid nanodiscs using single-particle cryo-electron microscopy. At a resolution of ~3 Å for the cytosolic domain and ~4 Å for the transmembrane domain, the structure determined in nanodiscs is similar to the previously determined X-ray structure. Our findings show that large differences in structure between detergent and lipid bilayer environments are unlikely, and enable us to propose possible structural mechanisms for C-type inactivation. | |||||||||
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_9024.map.gz | 132.6 MB | EMDB map data format | |
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Header (meta data) | emd-9024-v30.xml emd-9024.xml | 19.4 KB 19.4 KB | Display Display | EMDB header |
Images | emd_9024.png | 154.4 KB | ||
Masks | emd_9024_msk_1.map | 216 MB | Mask map | |
Filedesc metadata | emd-9024.cif.gz | 6.9 KB | ||
Others | emd_9024_additional.map.gz | 200.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9024 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9024 | HTTPS FTP |
-Validation report
Summary document | emd_9024_validation.pdf.gz | 437 KB | Display | EMDB validaton report |
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Full document | emd_9024_full_validation.pdf.gz | 436.6 KB | Display | |
Data in XML | emd_9024_validation.xml.gz | 7 KB | Display | |
Data in CIF | emd_9024_validation.cif.gz | 8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9024 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9024 | HTTPS FTP |
-Related structure data
Related structure data | 6ebkMC 9025C 9026C 6eblC 6ebmC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_9024.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Kv1.2-2.1 paddle chimera in lipid nanodiscs, entire protein | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.835 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_9024_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Kv1.2-2.1 paddle chimera in lipid nanodiscs, additional map
File | emd_9024_additional.map | ||||||||||||
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Annotation | Kv1.2-2.1 paddle chimera in lipid nanodiscs, additional map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in l...
Entire | Name: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs |
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Components |
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-Supramolecule #1: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in l...
Supramolecule | Name: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 385 KDa |
-Macromolecule #1: Voltage-gated potassium channel subunit beta-2
Macromolecule | Name: Voltage-gated potassium channel subunit beta-2 / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 37.339059 KDa |
Recombinant expression | Organism: Komagataella pastoris (fungus) |
Sequence | String: MVQFYRNLGK SGLRVSCLGL GTWVTFGGQI TDEMAEHLMT LAYDNGINLF DTAEVYAAGK AEVVLGNIIK KKGWRRSSLV ITTKIFWGG KAETERGLSR KHIIEGLKAS LERLQLEYVD VVFANRPDPN TPMEETVRAM THVINQGMAM YWGTSRWSSM E IMEAYSVA ...String: MVQFYRNLGK SGLRVSCLGL GTWVTFGGQI TDEMAEHLMT LAYDNGINLF DTAEVYAAGK AEVVLGNIIK KKGWRRSSLV ITTKIFWGG KAETERGLSR KHIIEGLKAS LERLQLEYVD VVFANRPDPN TPMEETVRAM THVINQGMAM YWGTSRWSSM E IMEAYSVA RQFNLIPPIC EQAEYHMFQR EKVEVQLPEL FHKIGVGAMT WSPLACGIVS GKYDSGIPPY SRASLKGYQW LK DKILSEE GRRQQAKLKE LQAIAERLGC TLPQLAIAWC LRNEGVSSVL LGASNAEQLM ENIGAIQVLP KLSSSIVHEI DSI LGNKPY SKKDYRS UniProtKB: Voltage-gated potassium channel subunit beta-2 |
-Macromolecule #2: Potassium voltage-gated channel subfamily A member 2,Potassium vo...
Macromolecule | Name: Potassium voltage-gated channel subfamily A member 2,Potassium voltage-gated channel subfamily B member 2 chimera type: protein_or_peptide / ID: 2 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 58.905828 KDa |
Recombinant expression | Organism: Komagataella pastoris (fungus) |
Sequence | String: MAHHHHHHHH ENLYFQGSMT VATGDPVDEA AAHPGHPQDT YDPEADHECC ERVVINISGL RFETQLKTLA QFPETLLGDP KKRMRYFDP LRNEYFFDRN RPSFDAILYY YQSGGRLRRP VNVPLDIFSE EIRFYELGEE AMEMFREDEG YIKEEERPLP E NEFQRQVW ...String: MAHHHHHHHH ENLYFQGSMT VATGDPVDEA AAHPGHPQDT YDPEADHECC ERVVINISGL RFETQLKTLA QFPETLLGDP KKRMRYFDP LRNEYFFDRN RPSFDAILYY YQSGGRLRRP VNVPLDIFSE EIRFYELGEE AMEMFREDEG YIKEEERPLP E NEFQRQVW LLFEYPESSG PARIIAIVSV MVILISIVSF CLETLPIFRD ENEDMHGGGV TFHTYSQSTI GYQQSTSFTD PF FIVETLC IIWFSFEFLV RFFACPSKAG FFTNIMNIID IVAIIPYYVT IFLTESNKSV LQFQNVRRVV QIFRIMRILR IFK LSRHSK GLQILGQTLK ASMRELGLLI FFLFIGVILF SSAVYFAEAD ERDSQFPSIP DAFWWAVVSM TTVGYGDMVP TTIG GKIVG SLCAIAGVLT IALPVPVIVS NFNYFYHRET EGEEQAQYLQ VTSCPKIPSS PDLKKSRSAS TISKSDYMEI QEGVN NSNE DFREENLKTA NCTLANTNYV NITKMLTDV UniProtKB: Potassium voltage-gated channel subfamily A member 2, Potassium voltage-gated channel subfamily B member 2, Potassium voltage-gated channel subfamily A member 2 |
-Macromolecule #3: NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE
Macromolecule | Name: NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / type: ligand / ID: 3 / Number of copies: 4 / Formula: NAP |
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Molecular weight | Theoretical: 743.405 Da |
Chemical component information | ChemComp-NAP: |
-Macromolecule #4: water
Macromolecule | Name: water / type: ligand / ID: 4 / Number of copies: 128 / Formula: HOH |
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Molecular weight | Theoretical: 18.015 Da |
Chemical component information | ChemComp-HOH: |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.7 mg/mL | ||||||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil, UltrAuFoil, R1.2/1.3 / Material: GOLD / Pretreatment - Type: PLASMA CLEANING | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 88 % / Chamber temperature: 277.15 K / Instrument: LEICA EM GP Details: A 3 microliter sample was applied to a plasma-cleaned grid and blotted for 10 seconds.. | ||||||||||||
Details | Kv1.2-2.1 paddle chimera in lipid nanodiscs |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Dimensions - Width: 7420 pixel / Digitization - Dimensions - Height: 7676 pixel / Digitization - Frames/image: 2-20 / Number grids imaged: 1 / Number real images: 3085 / Average exposure time: 15.2 sec. / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 29000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |