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- EMDB-9026: The voltage-activated Kv1.2-2.1 paddle chimera channel in lipid n... -

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Basic information

Entry
Database: EMDB / ID: EMD-9026
TitleThe voltage-activated Kv1.2-2.1 paddle chimera channel in lipid nanodiscs, transmembrane domain of subunit alpha
Map dataKv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
Sample
  • Complex: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
    • Protein or peptide: Potassium voltage-gated channel subfamily A member 2,Potassium voltage-gated channel subfamily B member 2 chimera
KeywordsMEMBRANE PROTEIN / transport protein / potassium channel / lipid nanodisc
Function / homology
Function and homology information


optic nerve structural organization / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / paranodal junction / potassium ion export across plasma membrane / regulation of circadian sleep/wake cycle, non-REM sleep / axon initial segment / delayed rectifier potassium channel activity / corpus callosum development ...optic nerve structural organization / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / paranodal junction / potassium ion export across plasma membrane / regulation of circadian sleep/wake cycle, non-REM sleep / axon initial segment / delayed rectifier potassium channel activity / corpus callosum development / voltage-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / outward rectifier potassium channel activity / juxtaparanode region of axon / optic nerve development / neuronal cell body membrane / regulation of dopamine secretion / voltage-gated potassium channel activity / lamellipodium membrane / kinesin binding / neuronal action potential / calyx of Held / voltage-gated potassium channel complex / axon terminus / sensory perception of pain / potassium ion transmembrane transport / protein localization to plasma membrane / protein homooligomerization / potassium ion transport / cerebral cortex development / lamellipodium / presynaptic membrane / perikaryon / postsynaptic membrane / transmembrane transporter binding / endosome / protein heterodimerization activity / axon / dendrite / neuronal cell body / glutamatergic synapse / endoplasmic reticulum membrane / plasma membrane
Similarity search - Function
Potassium channel, voltage dependent, Kv2.2 / Potassium channel, voltage dependent, Kv2 / Potassium channel, voltage dependent, Kv1.2 / Kv2 voltage-gated K+ channel / Potassium channel, voltage dependent, Kv1 / Potassium channel, voltage dependent, Kv / Potassium channel tetramerisation-type BTB domain / BTB/POZ domain / Broad-Complex, Tramtrack and Bric a brac / BTB/POZ domain ...Potassium channel, voltage dependent, Kv2.2 / Potassium channel, voltage dependent, Kv2 / Potassium channel, voltage dependent, Kv1.2 / Kv2 voltage-gated K+ channel / Potassium channel, voltage dependent, Kv1 / Potassium channel, voltage dependent, Kv / Potassium channel tetramerisation-type BTB domain / BTB/POZ domain / Broad-Complex, Tramtrack and Bric a brac / BTB/POZ domain / Voltage-dependent channel domain superfamily / SKP1/BTB/POZ domain superfamily / Ion transport domain / Ion transport protein
Similarity search - Domain/homology
Potassium voltage-gated channel subfamily A member 2 / Potassium voltage-gated channel subfamily B member 2
Similarity search - Component
Biological speciesRattus norvegicus (Norway rat)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.0 Å
AuthorsMatthies D / Bae C
CitationJournal: 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
DepositionAug 6, 2018-
Header (metadata) releaseAug 22, 2018-
Map releaseAug 22, 2018-
UpdateMar 13, 2024-
Current statusMar 13, 2024Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6ebm
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
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Supplemental images

Downloads & links

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Map

FileDownload / File: emd_9026.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationKv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
Voxel sizeX=Y=Z: 0.835 Å
Density
Contour LevelBy AUTHOR: 0.02 / Movie #1: 0.02
Minimum - Maximum-0.04445777 - 0.092283696
Average (Standard dev.)-0.000012591698 (±0.0015055928)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 320.63998 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.8350.8350.835
M x/y/z384384384
origin x/y/z0.0000.0000.000
length x/y/z320.640320.640320.640
α/β/γ90.00090.00090.000
start NX/NY/NZ-34-26-36
NX/NY/NZ528549
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS384384384
D min/max/mean-0.0440.092-0.000

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Supplemental data

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Mask #1

Fileemd_9026_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in l...

EntireName: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
Components
  • Complex: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
    • Protein or peptide: Potassium voltage-gated channel subfamily A member 2,Potassium voltage-gated channel subfamily B member 2 chimera

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Supramolecule #1: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in l...

SupramoleculeName: Voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane domain
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Rattus norvegicus (Norway rat)
Molecular weightTheoretical: 385 KDa

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Macromolecule #1: Potassium voltage-gated channel subfamily A member 2,Potassium vo...

MacromoleculeName: Potassium voltage-gated channel subfamily A member 2,Potassium voltage-gated channel subfamily B member 2 chimera
type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Rattus norvegicus (Norway rat)
Molecular weightTheoretical: 58.905828 KDa
Recombinant expressionOrganism: Komagataella pastoris (fungus)
SequenceString: 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

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.7 mg/mL
BufferpH: 7.5
Component:
ConcentrationFormulaName
20.0 mMHEPES-KOHHEPES-KOH
150.0 mMKClpotassium chloride
2.0 mMTCEPTCEP
GridModel: Quantifoil, UltrAuFoil, R1.2/1.3 / Material: GOLD / Pretreatment - Type: PLASMA CLEANING
VitrificationCryogen 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..
DetailsKv1.2-2.1 paddle chimera in lipid nanodiscs, transmembrane alpha subunits

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 29000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm 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
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 281021
Startup modelType of model: OTHER
Details: An initial model was created from negatively stained single particle electron microscopy.
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final 3D classificationNumber classes: 2 / Software - Name: RELION (ver. 2.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C4 (4 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 4.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 2.1) / Number images used: 65745

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Atomic model buiding 1

Initial modelPDB ID:

Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: FLEXIBLE FIT / Overall B value: 123
Output model

PDB-6ebm:
The voltage-activated Kv1.2-2.1 paddle chimera channel in lipid nanodiscs, transmembrane domain of subunit alpha

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