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- EMDB-31016: CryoEM structure of human Kv4.2-DPP6S-KChIP1 complex, transmembra... -

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Basic information

Entry
Database: EMDB / ID: EMD-31016
TitleCryoEM structure of human Kv4.2-DPP6S-KChIP1 complex, transmembrane and intracellular region
Map datamap I
Sample
  • Cell: human Kv4.2-DPP6S-KChIP1 complex
    • Protein or peptide: Kv channel-interacting protein 1
    • Protein or peptide: Potassium voltage-gated channel subfamily D member 2
    • Protein or peptide: Dipeptidyl aminopeptidase-like protein 6
    • Protein or peptide: Kv channel-interacting protein 1
    • Protein or peptide: Dipeptidyl aminopeptidase-like protein 6
Keywordsmembrane protein
Function / homology
Function and homology information


Kv4.3-KChIP1 channel complex / Kv4.2-KChIP2 channel complex / A-type (transient outward) potassium channel activity / Phase 1 - inactivation of fast Na+ channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium ion export across plasma membrane / membrane repolarization / Voltage gated Potassium channels / regulation of potassium ion transmembrane transport / anchoring junction ...Kv4.3-KChIP1 channel complex / Kv4.2-KChIP2 channel complex / A-type (transient outward) potassium channel activity / Phase 1 - inactivation of fast Na+ channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium ion export across plasma membrane / membrane repolarization / Voltage gated Potassium channels / regulation of potassium ion transmembrane transport / anchoring junction / postsynaptic specialization membrane / regulation of heart contraction / action potential / neuronal cell body membrane / plasma membrane raft / locomotor rhythm / voltage-gated potassium channel activity / potassium channel activity / neuronal action potential / potassium channel regulator activity / voltage-gated potassium channel complex / GABA-ergic synapse / muscle contraction / potassium ion transmembrane transport / sensory perception of pain / serine-type peptidase activity / protein localization to plasma membrane / protein homooligomerization / cytoplasmic side of plasma membrane / cellular response to hypoxia / chemical synaptic transmission / perikaryon / postsynaptic membrane / transmembrane transporter binding / dendritic spine / neuronal cell body / glutamatergic synapse / dendrite / calcium ion binding / synapse / proteolysis / membrane / metal ion binding / plasma membrane / cytoplasm
Similarity search - Function
Potassium channel, voltage dependent, Kv4.2 / Potassium channel, voltage dependent, Kv4 / Potassium channel, voltage dependent, Kv4, C-terminal / Domain of unknown function (DUF3399) / Shal-type voltage-gated potassium channels, N-terminal / Shal-type voltage-gated potassium channels, N-terminal / Recoverin family / Potassium channel, voltage dependent, Kv / Potassium channel tetramerisation-type BTB domain / BTB/POZ domain ...Potassium channel, voltage dependent, Kv4.2 / Potassium channel, voltage dependent, Kv4 / Potassium channel, voltage dependent, Kv4, C-terminal / Domain of unknown function (DUF3399) / Shal-type voltage-gated potassium channels, N-terminal / Shal-type voltage-gated potassium channels, N-terminal / Recoverin family / Potassium channel, voltage dependent, Kv / Potassium channel tetramerisation-type BTB domain / BTB/POZ domain / : / Dipeptidylpeptidase IV, N-terminal domain / Dipeptidyl peptidase IV (DPP IV) N-terminal region / EF-hand domain pair / Peptidase S9, prolyl oligopeptidase, catalytic domain / Prolyl oligopeptidase family / Broad-Complex, Tramtrack and Bric a brac / BTB/POZ domain / Voltage-dependent channel domain superfamily / SKP1/BTB/POZ domain superfamily / EF-hand domain pair / EF-hand, calcium binding motif / EF-Hand 1, calcium-binding site / EF-hand calcium-binding domain. / EF-hand calcium-binding domain profile. / EF-hand domain / Ion transport domain / Ion transport protein / EF-hand domain pair / Alpha/Beta hydrolase fold
Similarity search - Domain/homology
A-type potassium channel modulatory protein DPP6 / A-type potassium channel modulatory protein KCNIP1 / A-type voltage-gated potassium channel KCND2
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.9 Å
AuthorsKise Y / Nureki O
Funding support Japan, 1 items
OrganizationGrant numberCountry
Japan Society for the Promotion of Science (JSPS) Japan
CitationJournal: Nature / Year: 2021
Title: Structural basis of gating modulation of Kv4 channel complexes.
Authors: Yoshiaki Kise / Go Kasuya / Hiroyuki H Okamoto / Daichi Yamanouchi / Kan Kobayashi / Tsukasa Kusakizako / Tomohiro Nishizawa / Koichi Nakajo / Osamu Nureki /
Abstract: Modulation of voltage-gated potassium (Kv) channels by auxiliary subunits is central to the physiological function of channels in the brain and heart. Native Kv4 tetrameric channels form ...Modulation of voltage-gated potassium (Kv) channels by auxiliary subunits is central to the physiological function of channels in the brain and heart. Native Kv4 tetrameric channels form macromolecular ternary complexes with two auxiliary β-subunits-intracellular Kv channel-interacting proteins (KChIPs) and transmembrane dipeptidyl peptidase-related proteins (DPPs)-to evoke rapidly activating and inactivating A-type currents, which prevent the backpropagation of action potentials. However, the modulatory mechanisms of Kv4 channel complexes remain largely unknown. Here we report cryo-electron microscopy structures of the Kv4.2-DPP6S-KChIP1 dodecamer complex, the Kv4.2-KChIP1 and Kv4.2-DPP6S octamer complexes, and Kv4.2 alone. The structure of the Kv4.2-KChIP1 complex reveals that the intracellular N terminus of Kv4.2 interacts with its C terminus that extends from the S6 gating helix of the neighbouring Kv4.2 subunit. KChIP1 captures both the N and the C terminus of Kv4.2. In consequence, KChIP1 would prevent N-type inactivation and stabilize the S6 conformation to modulate gating of the S6 helices within the tetramer. By contrast, unlike the reported auxiliary subunits of voltage-gated channel complexes, DPP6S interacts with the S1 and S2 helices of the Kv4.2 voltage-sensing domain, which suggests that DPP6S stabilizes the conformation of the S1-S2 helices. DPP6S may therefore accelerate the voltage-dependent movement of the S4 helices. KChIP1 and DPP6S do not directly interact with each other in the Kv4.2-KChIP1-DPP6S ternary complex. Thus, our data suggest that two distinct modes of modulation contribute in an additive manner to evoke A-type currents from the native Kv4 macromolecular complex.
History
DepositionMar 1, 2021-
Header (metadata) releaseOct 13, 2021-
Map releaseOct 13, 2021-
UpdateJun 5, 2024-
Current statusJun 5, 2024Processing site: PDBj / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0163
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.0163
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-7e8e
  • Surface level: 0.0163
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_31016.map.gz / Format: CCP4 / Size: 101 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationmap I
Projections & slices

Image control

Size
Brightness
Contrast
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AxesZ (Sec.)Y (Row.)X (Col.)
1 Å/pix.
x 298 pix.
= 298.799 Å
1 Å/pix.
x 298 pix.
= 298.799 Å
1 Å/pix.
x 298 pix.
= 298.799 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.00268 Å
Density
Contour LevelBy AUTHOR: 0.0163 / Movie #1: 0.0163
Minimum - Maximum-0.04339206 - 0.08375512
Average (Standard dev.)0.00024129437 (±0.0020418346)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions298298298
Spacing298298298
CellA=B=C: 298.7986 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.00268120805371.00268120805371.0026812080537
M x/y/z298298298
origin x/y/z0.0000.0000.000
length x/y/z298.799298.799298.799
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS298298298
D min/max/mean-0.0430.0840.000

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Supplemental data

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Mask #1

Fileemd_31016_msk_1.map
Projections & Slices
AxesZYX

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Half map: #1

Fileemd_31016_half_map_1.map
Projections & Slices
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Half map: #2

Fileemd_31016_half_map_2.map
Projections & Slices
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Sample components

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Entire : human Kv4.2-DPP6S-KChIP1 complex

EntireName: human Kv4.2-DPP6S-KChIP1 complex
Components
  • Cell: human Kv4.2-DPP6S-KChIP1 complex
    • Protein or peptide: Kv channel-interacting protein 1
    • Protein or peptide: Potassium voltage-gated channel subfamily D member 2
    • Protein or peptide: Dipeptidyl aminopeptidase-like protein 6
    • Protein or peptide: Kv channel-interacting protein 1
    • Protein or peptide: Dipeptidyl aminopeptidase-like protein 6

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Supramolecule #1: human Kv4.2-DPP6S-KChIP1 complex

SupramoleculeName: human Kv4.2-DPP6S-KChIP1 complex / type: cell / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Kv channel-interacting protein 1

MacromoleculeName: Kv channel-interacting protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 21.242941 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
PEGLEQLEAQ TNFTKRELQV LYRGFKNECP SGVVNEDTFK QIYAQFFPHG DASTYAHYLF NAFDTTQTGS VKFEDFVTAL SILLRGTVH EKLRWTFNLY DINKDGYINK EEMMDIVKAI YDMMGKYTYP VLKEDTPRQH VDVFFQKMDK NKDGIVTLDE F LESCQEDD NIMRSLQLFQ NVM

UniProtKB: A-type potassium channel modulatory protein KCNIP1

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Macromolecule #2: Potassium voltage-gated channel subfamily D member 2

MacromoleculeName: Potassium voltage-gated channel subfamily D member 2 / type: protein_or_peptide / ID: 2 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 55.755738 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: AAGVAAWLPF ARAAAIGWMP VASGPMPAPP RQERKRTQDA LIVLNVSGTR FQTWQDTLER YPDTLLGSSE RDFFYHPETQ QYFFDRDPD IFRHILNFYR TGKLHYPRHE CISAYDEELA FFGLIPEIIG DCCYEEYKDR RRENAERLQD DADTDTAGES A LPTMTARQ ...String:
AAGVAAWLPF ARAAAIGWMP VASGPMPAPP RQERKRTQDA LIVLNVSGTR FQTWQDTLER YPDTLLGSSE RDFFYHPETQ QYFFDRDPD IFRHILNFYR TGKLHYPRHE CISAYDEELA FFGLIPEIIG DCCYEEYKDR RRENAERLQD DADTDTAGES A LPTMTARQ RVWRAFENPH TSTMALVFYY VTGFFIAVSV IANVVETVPC GSSPGHIKEL PCGERYAVAF FCLDTACVMI FT VEYLLRL AAAPSRYRFV RSVMSIIDVV AILPYYIGLV MTDNEDVSGA FVTLRVFRVF RIFKFSRHSQ GLRILGYTLK SCA SELGFL LFSLTMAIII FATVMFYAEK GSSASKFTSI PAAFWYTIVT MTTLGYGDMV PKTIAGKIFG SICSLSGVLV IALP VPVIV SNFSRIYHQN QRADKRRAQK KARLARIRAA KSGSANAYMQ SKRSGLLSNQ LQSSEDEQAF VSKSGSSFET QHHHL LHCL EKTTNHEFVD

UniProtKB: A-type voltage-gated potassium channel KCND2

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Macromolecule #3: Dipeptidyl aminopeptidase-like protein 6

MacromoleculeName: Dipeptidyl aminopeptidase-like protein 6 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 3.032829 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
AAAAKGIAIA LLVILVICSL IVTSVILLTP A

UniProtKB: A-type potassium channel modulatory protein DPP6

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Macromolecule #4: Kv channel-interacting protein 1

MacromoleculeName: Kv channel-interacting protein 1 / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 21.145826 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
EGLEQLEAQT NFTKRELQVL YRGFKNECPS GVVNEDTFKQ IYAQFFPHGD ASTYAHYLFN AFDTTQTGSV KFEDFVTALS ILLRGTVHE KLRWTFNLYD INKDGYINKE EMMDIVKAIY DMMGKYTYPV LKEDTPRQHV DVFFQKMDKN KDGIVTLDEF L ESCQEDDN IMRSLQLFQN VM

UniProtKB: A-type potassium channel modulatory protein KCNIP1

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Macromolecule #5: Dipeptidyl aminopeptidase-like protein 6

MacromoleculeName: Dipeptidyl aminopeptidase-like protein 6 / type: protein_or_peptide / ID: 5 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 2.819595 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
AKGIAIALLV ILVICSLIVT SVILLTPA

UniProtKB: A-type potassium channel modulatory protein DPP6

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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

BufferpH: 8
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 48.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 139524
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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