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- EMDB-20294: Single particle cryo-EM structure of the voltage-gated K+ channel... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-20294 | ||||||||||||
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Title | Single particle cryo-EM structure of the voltage-gated K+ channel Eag1 3-13 deletion mutant bound to calmodulin (conformation 2) | ||||||||||||
![]() | Sharpened map for Eag1 3-13/CaM conformation 2 | ||||||||||||
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![]() | Voltage-gated potassium channel / ion channel / calmodulin / TRANSPORT PROTEIN-CALCIUM BINDING PROTEIN complex | ||||||||||||
Function / homology | ![]() Voltage gated Potassium channels / potassium channel complex / regulation of presynaptic cytosolic calcium ion concentration / voltage-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / delayed rectifier potassium channel activity / CaM pathway / parallel fiber to Purkinje cell synapse / Cam-PDE 1 activation / Sodium/Calcium exchangers / Calmodulin induced events ...Voltage gated Potassium channels / potassium channel complex / regulation of presynaptic cytosolic calcium ion concentration / voltage-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / delayed rectifier potassium channel activity / CaM pathway / parallel fiber to Purkinje cell synapse / Cam-PDE 1 activation / Sodium/Calcium exchangers / Calmodulin induced events / nuclear inner membrane / Reduction of cytosolic Ca++ levels / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / Activation of Ca-permeable Kainate Receptor / Loss of phosphorylation of MECP2 at T308 / CREB1 phosphorylation through the activation of Adenylate Cyclase / PKA activation / negative regulation of high voltage-gated calcium channel activity / CaMK IV-mediated phosphorylation of CREB / Glycogen breakdown (glycogenolysis) / phosphatidylinositol bisphosphate binding / positive regulation of cyclic-nucleotide phosphodiesterase activity / organelle localization by membrane tethering / negative regulation of calcium ion export across plasma membrane / regulation of synaptic vesicle exocytosis / CLEC7A (Dectin-1) induces NFAT activation / regulation of cardiac muscle cell action potential / mitochondrion-endoplasmic reticulum membrane tethering / autophagosome membrane docking / Activation of RAC1 downstream of NMDARs / positive regulation of ryanodine-sensitive calcium-release channel activity / regulation of cell communication by electrical coupling involved in cardiac conduction / Negative regulation of NMDA receptor-mediated neuronal transmission / startle response / negative regulation of peptidyl-threonine phosphorylation / Synthesis of IP3 and IP4 in the cytosol / Unblocking of NMDA receptors, glutamate binding and activation / Phase 0 - rapid depolarisation / protein phosphatase activator activity / RHO GTPases activate PAKs / positive regulation of phosphoprotein phosphatase activity / Ion transport by P-type ATPases / Long-term potentiation / Uptake and function of anthrax toxins / Calcineurin activates NFAT / Regulation of MECP2 expression and activity / catalytic complex / DARPP-32 events / detection of calcium ion / regulation of cardiac muscle contraction / axolemma / negative regulation of ryanodine-sensitive calcium-release channel activity / Smooth Muscle Contraction / calcium channel inhibitor activity / RHO GTPases activate IQGAPs / cellular response to interferon-beta / regulation of cardiac muscle contraction by regulation of the release of sequestered calcium ion / Protein methylation / eNOS activation / Activation of AMPK downstream of NMDARs / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation / positive regulation of protein dephosphorylation / Ion homeostasis / regulation of calcium-mediated signaling / regulation of ryanodine-sensitive calcium-release channel activity / titin binding / monoatomic ion transmembrane transport / positive regulation of protein autophosphorylation / potassium ion transmembrane transport / voltage-gated potassium channel complex / sperm midpiece / 14-3-3 protein binding / calcium channel complex / cellular response to calcium ion / substantia nigra development / adenylate cyclase activator activity / Ras activation upon Ca2+ influx through NMDA receptor / regulation of heart rate / sarcomere / protein serine/threonine kinase activator activity / FCERI mediated Ca+2 mobilization / FCGR3A-mediated IL10 synthesis / VEGFR2 mediated vascular permeability / regulation of cytokinesis / Antigen activates B Cell Receptor (BCR) leading to generation of second messengers / positive regulation of peptidyl-threonine phosphorylation / VEGFR2 mediated cell proliferation / Translocation of SLC2A4 (GLUT4) to the plasma membrane / regulation of membrane potential / postsynaptic density membrane / positive regulation of receptor signaling pathway via JAK-STAT / RAF activation / Transcriptional activation of mitochondrial biogenesis / positive regulation of protein serine/threonine kinase activity / spindle microtubule / Stimuli-sensing channels / spindle pole / cellular response to type II interferon / response to calcium ion Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.0 Å | ||||||||||||
![]() | Whicher JR / MacKinnon R | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Regulation of Eag1 gating by its intracellular domains. Authors: Jonathan R Whicher / Roderick MacKinnon / ![]() Abstract: Voltage-gated potassium channels (Ks) are gated by transmembrane voltage sensors (VS) that move in response to changes in membrane voltage. K10.1 or Eag1 also has three intracellular domains: PAS, C- ...Voltage-gated potassium channels (Ks) are gated by transmembrane voltage sensors (VS) that move in response to changes in membrane voltage. K10.1 or Eag1 also has three intracellular domains: PAS, C-linker, and CNBHD. We demonstrate that the Eag1 intracellular domains are not required for voltage-dependent gating but likely interact with the VS to modulate gating. We identified specific interactions between the PAS, CNBHD, and VS that modulate voltage-dependent gating and provide evidence that VS movement destabilizes these interactions to promote channel opening. Additionally, mutation of these interactions renders Eag1 insensitive to calmodulin inhibition. The structure of the calmodulin insensitive mutant in a pre-open conformation suggests that channel opening may occur through a rotation of the intracellular domains and calmodulin may prevent this rotation by stabilizing interactions between the VS and intracellular domains. Intracellular domains likely play a similar modulatory role in voltage-dependent gating of the related K11-12 channels. | ||||||||||||
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 | ![]() | 116.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 16 KB 16 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.2 KB | Display | ![]() |
Images | ![]() | 196.3 KB | ||
Filedesc metadata | ![]() | 6.1 KB | ||
Others | ![]() | 92.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 622.7 KB | Display | ![]() |
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Full document | ![]() | 622.3 KB | Display | |
Data in XML | ![]() | 10.6 KB | Display | |
Data in CIF | ![]() | 14.5 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6pbxMC ![]() 6pbyC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Sharpened map for Eag1 3-13/CaM conformation 2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Unsharpened map for Eag1 3-13/CaM conformation 2
File | emd_20294_additional.map | ||||||||||||
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Annotation | Unsharpened map for Eag1 3-13/CaM conformation 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Voltage-gated potassium channel Eag1 3-13 deletion mutant bound t...
Entire | Name: Voltage-gated potassium channel Eag1 3-13 deletion mutant bound to calmodulin |
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Components |
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-Supramolecule #1: Voltage-gated potassium channel Eag1 3-13 deletion mutant bound t...
Supramolecule | Name: Voltage-gated potassium channel Eag1 3-13 deletion mutant bound to calmodulin type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Supramolecule #2: Voltage-gated potassium channel Eag1 3-13 deletion mutant
Supramolecule | Name: Voltage-gated potassium channel Eag1 3-13 deletion mutant type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
-Supramolecule #3: calmodulin
Supramolecule | Name: calmodulin / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Potassium voltage-gated channel subfamily H member 1
Macromolecule | Name: Potassium voltage-gated channel subfamily H member 1 / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 96.26143 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MAQNTFLENI VRRSNDTNFV LGNAQIVDWP IVYSNDGFCK LSGYHRAEVM QKSSACSFMY GELTDKDTVE KVRQTFENYE MNSFEILMY KKNRTPVWFF VKIAPIRNEQ DKVVLFLCTF SDITAFKQPI EDDSCKGWGK FARLTRALTS SRGVLQQLAP S VQKGENVH ...String: MAQNTFLENI VRRSNDTNFV LGNAQIVDWP IVYSNDGFCK LSGYHRAEVM QKSSACSFMY GELTDKDTVE KVRQTFENYE MNSFEILMY KKNRTPVWFF VKIAPIRNEQ DKVVLFLCTF SDITAFKQPI EDDSCKGWGK FARLTRALTS SRGVLQQLAP S VQKGENVH KHSRLAEVLQ LGSDILPQYK QEAPKTPPHI ILHYCVFKTT WDWIILILTF YTAILVPYNV SFKTRQNNVA WL VVDSIVD VIFLVDIVLN FHTTFVGPAG EVISDPKLIR MNYLKTWFVI DLLSCLPYDV INAFENVDEG ISSLFSSLKV VRL LRLGRV ARKLDHYIEY GAAVLVLLVC VFGLAAHWMA CIWYSIGDYE IFDEDTKTIR NNSWLYQLAL DIGTPYQFNG SGSG KWEGG PSKNSVYISS LYFTMTSLTS VGFGNIAPST DIEKIFAVAI MMIGSLLYAT IFGNVTTIFQ QMYANTNRYH EMLNS VRDF LKLYQVPKGL SERVMDYIVS TWSMSRGIDT EKVLQICPKD MRADICVHLN RKVFKEHPAF RLASDGCLRA LAMEFQ TVH CAPGDLIYHA GESVDSLCFV VSGSLEVIQD DEVVAILGKG DVFGDVFWKE ATLAQSCANV RALTYCDLHV IKRDALQ KV LEFYTAFSHS FSRNLILTYN LRKRIVFRKI SDVKREEEER MKRKNEAPLI LPPDHPVRRL FQRFRQQKEA RLAAERGG R DLDDLDVEKG NALTDHTSAN HSLVKASVVT VRESPATPVS FYPIPEQTLQ ATVLEVKHEL KEDIKALNAK MTSIEKQLS EILRILMSRG SSQSPQDTCE VSRPQSPESD RDIFGASSNS LEVLFQ UniProtKB: Potassium voltage-gated channel subfamily H member 1, Potassium voltage-gated channel subfamily H member 1 |
-Macromolecule #2: Calmodulin-1
Macromolecule | Name: Calmodulin-1 / type: protein_or_peptide / ID: 2 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 16.852545 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MADQLTEEQI AEFKEAFSLF DKDGDGTITT KELGTVMRSL GQNPTEAELQ DMINEVDADG NGTIDFPEFL TMMARKMKDT DSEEEIREA FRVFDKDGNG YISAAELRHV MTNLGEKLTD EEVDEMIREA DIDGDGQVNY EEFVQMMTAK UniProtKB: Calmodulin-1 |
-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: 8 |
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Grid | Details: unspecified |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 1.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |