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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-0439 | |||||||||
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Title | human BK channel reconstituted into liposomes | |||||||||
![]() | human BK channel reconstituted into liposomes | |||||||||
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![]() | the large conductance voltage- and calcium-activated potassium (BK) channel / Ca2+ sensor / gating ring / random spherically constrained (RSC) single-particle cryo-EM / MEMBRANE PROTEIN | |||||||||
Function / homology | ![]() Acetylcholine inhibits contraction of outer hair cells / micturition / Ca2+ activated K+ channels / large conductance calcium-activated potassium channel activity / response to carbon monoxide / calcium-activated potassium channel activity / negative regulation of cell volume / smooth muscle contraction involved in micturition / intracellular potassium ion homeostasis / Sensory processing of sound by inner hair cells of the cochlea ...Acetylcholine inhibits contraction of outer hair cells / micturition / Ca2+ activated K+ channels / large conductance calcium-activated potassium channel activity / response to carbon monoxide / calcium-activated potassium channel activity / negative regulation of cell volume / smooth muscle contraction involved in micturition / intracellular potassium ion homeostasis / Sensory processing of sound by inner hair cells of the cochlea / response to osmotic stress / cGMP effects / voltage-gated potassium channel activity / voltage-gated potassium channel complex / potassium ion transmembrane transport / potassium ion transport / regulation of membrane potential / caveola / response to calcium ion / vasodilation / actin binding / postsynaptic membrane / response to hypoxia / apical plasma membrane / positive regulation of apoptotic process / identical protein binding / membrane / metal ion binding / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | |||||||||
![]() | Wang L / Tonggu L | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Broken symmetry in the human BK channel Authors: Tonggu L / Wang L | |||||||||
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 | ![]() | 59.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 16.6 KB 16.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.1 KB | Display | ![]() |
Images | ![]() | 154.2 KB | ||
Filedesc metadata | ![]() | 7.2 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 649 KB | Display | ![]() |
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Full document | ![]() | 648.6 KB | Display | |
Data in XML | ![]() | 11.6 KB | Display | |
Data in CIF | ![]() | 15.5 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6nd0MC M: atomic model generated by this map C: citing same article ( |
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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 | human BK channel reconstituted into liposomes | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.056 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : human BK channel reconstituted into liposomes
Entire | Name: human BK channel reconstituted into liposomes |
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Components |
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-Supramolecule #1: human BK channel reconstituted into liposomes
Supramolecule | Name: human BK channel reconstituted into liposomes / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 500 KDa |
-Macromolecule #1: Calcium-activated potassium channel subunit alpha-1
Macromolecule | Name: Calcium-activated potassium channel subunit alpha-1 / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 99.256867 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: LKTSNSIKLV NLLSIFISTW LTAAGFIHLV ENSGDPWENF QNNQALTYWE CVYLLMVTMS TVGYGDVYAK TTLGRLFMVF FILGGLAMF ASYVPEIIEL IGNRKKYGGS YSAVSGRKHI VVCGHITLES VSNFLKDFLH KDRDDVNVEI VFLHNISPNL E LEALFKRH ...String: LKTSNSIKLV NLLSIFISTW LTAAGFIHLV ENSGDPWENF QNNQALTYWE CVYLLMVTMS TVGYGDVYAK TTLGRLFMVF FILGGLAMF ASYVPEIIEL IGNRKKYGGS YSAVSGRKHI VVCGHITLES VSNFLKDFLH KDRDDVNVEI VFLHNISPNL E LEALFKRH FTQVEFYQGS VLNPHDLARV KIESADACLI LANKYCADPD AEDASNIMRV ISIKNYHPKI RIITQMLQYH NK AHLLNIP SWNWKEGDDA ICLAELKLGF IAQSCLAQGL STMLANLFSM RSFIKIEEDT WQKYYLEGVS NEMYTEYLSS AFV GLSFPT VCELCFVKLK LLMIAIEYKS ANRESRILIN PGNHLKIQEG TLGFFIASDA KEVKRAFFYC KACHDDITDP KRIK KCGCK RPKMSIYKRM RRACCFDCGR SERDCSCMSG RVRGNVDTLE RAFPLSSVSV NDCSTSFRAF EDEQPSTLSP KKKQR NGGM RNSPNTSPKL MRHDPLLIPG NDQIDNMDSN VKKYDSTGMF HWCAPKEIEK VILTRSEAAM TVLSGHVVVC IFGDVS SAL IGLRNLVMPL RASNFHYHEL KHIVFVGSIE YLKREWETLH NFPKVSILPG TPLSRADLRA VNINLCDMCV ILSANQN NI DDTSLQDKEC ILASLNIKSM QFDDSIGVLQ ANSQGFTPPG MDRSSPDNSP VHGMLRQPSI TTGVNIPIIT ELVNDTNV Q FLDQDDDDDP DTELYLTQPF ACGTAFAVSV LDSLMSATYF NDNILTLIRT LVTGGATPEL EALIAEENAL RGGYSTPQT LANRDRCRVA QLALLDGPFA DLGDGGCYGD LFCKALKTYN MLCFGIYRLR DAHLSTPSQC TKRYVITNPP YEFELVPTDL IFCL UniProtKB: Calcium-activated potassium channel subunit alpha-1 |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1.0 mg/mL | ||||||||||||
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Buffer | pH: 7.3 Component:
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Grid | Model: C-flat-2/2 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 20 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 20 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.133 kPa | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV Details: BK proteoliposomes were applied onto a glow-discharged perforated carbon grid (CFlat R2/2, EMS), and incubated for 5-10 min In an FEI Vitrobot Mark IV (FEI). After incubation, the sample was ...Details: BK proteoliposomes were applied onto a glow-discharged perforated carbon grid (CFlat R2/2, EMS), and incubated for 5-10 min In an FEI Vitrobot Mark IV (FEI). After incubation, the sample was blotted from the side and buffer containing 10 mM HEPES, pH 7.3, 75 mM KCl, and 1 mM EDTA was applied onto the TEM grid to further swell proteoliposomes. Then grids were blotted for 7 s with force 0 and fast frozen in liquid ethane cooled by liquid nitrogen.. | ||||||||||||
Details | Swelled BK proteoliposomes yielded a concentration of 2 mM lipid and 1mg/ml protein. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Temperature | Min: 90.8 K / Max: 103.5 K |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Details | The Coma free alignment was performed using Auto-CTF software from FEI. |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-43 / Number grids imaged: 3 / Number real images: 3089 / Average exposure time: 8.6 sec. / Average electron dose: 60.0 e/Å2 Details: Images were recorded in an automated fashion on a Gatan K2 Summit (Gatan) detector set to super-resolution mode with a super-resolution pixel size of 0.525 A using Leginon. The dose rate on ...Details: Images were recorded in an automated fashion on a Gatan K2 Summit (Gatan) detector set to super-resolution mode with a super-resolution pixel size of 0.525 A using Leginon. The dose rate on the camera was set to be ~8 counts per physical pixel per second. The total exposure time was 8.6 s (0.2 s/frame), leading to a total accumulate dose of 60 electrons per A2 on the specimen. |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 4.0 µm / Nominal defocus min: 0.852 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
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Output model | ![]() PDB-6nd0: |