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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-9323 | |||||||||
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Title | BEST1 open state W287F mutant, calcium-bound | |||||||||
![]() | BEST1 open state W287F mutant, calcium-bound | |||||||||
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Function / homology | Bestrophin-1 / Stimuli-sensing channels / Bestrophin / Bestrophin/UPF0187 / Bestrophin, RFP-TM, chloride channel / chloride channel activity / membrane / metal ion binding / Bestrophin 1![]() | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.7 Å | |||||||||
![]() | Miller AN / Vaisey G / Long SB | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Molecular mechanisms of gating in the calcium-activated chloride channel bestrophin. Authors: Alexandria N Miller / George Vaisey / Stephen B Long / ![]() Abstract: Bestrophin (BEST1-4) ligand-gated chloride (Cl) channels are activated by calcium (Ca). Mutation of BEST1 causes retinal disease. Partly because bestrophin channels have no sequence or structural ...Bestrophin (BEST1-4) ligand-gated chloride (Cl) channels are activated by calcium (Ca). Mutation of BEST1 causes retinal disease. Partly because bestrophin channels have no sequence or structural similarity to other ion channels, the molecular mechanisms underlying gating are unknown. Here, we present a series of cryo-electron microscopy structures of chicken BEST1, determined at 3.1 Å resolution or better, that represent the channel's principal gating states. Unlike other channels, opening of the pore is due to the repositioning of tethered pore-lining helices within a surrounding protein shell that dramatically widens a neck of the pore through a concertina of amino acid rearrangements. The neck serves as both the activation and the inactivation gate. Ca binding instigates opening of the neck through allosteric means whereas inactivation peptide binding induces closing. An aperture within the otherwise wide pore controls anion permeability. The studies define a new molecular paradigm for gating among ligand-gated ion 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 | ![]() | 12 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 12.9 KB 12.9 KB | Display Display | ![]() |
Images | ![]() | 181.5 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 472.5 KB | Display | ![]() |
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Full document | ![]() | 472.1 KB | Display | |
Data in XML | ![]() | 5.5 KB | Display | |
Data in CIF | ![]() | 6.2 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6n25MC ![]() 9321C ![]() 9322C ![]() 9324C ![]() 9325C ![]() 9326C ![]() 6n23C ![]() 6n24C ![]() 6n26C ![]() 6n27C ![]() 6n28C 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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Map
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Annotation | BEST1 open state W287F mutant, calcium-bound | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.088 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 : BEST1 open state W287F mutant, calcium-bound
Entire | Name: BEST1 open state W287F mutant, calcium-bound |
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Components |
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-Supramolecule #1: BEST1 open state W287F mutant, calcium-bound
Supramolecule | Name: BEST1 open state W287F mutant, calcium-bound / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() |
-Macromolecule #1: Bestrophin homolog
Macromolecule | Name: Bestrophin homolog / type: protein_or_peptide / ID: 1 / Number of copies: 5 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 40.722688 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: TVTYTNRVAD ARLGTFSQLL LQWKGSIYKL LYSEFLIFIS LYFAISLVYR LILSESQRLM FEKLALYCNS YAELIPVSFV LGFYVSLVV SRWWAQYESI PWPDRIMNLV SCNVDGEDEY GRLLRRTLMR YSNLCSVLIL RSVSTAVYKR FPSMEHVVRA G LMTPEEHK ...String: TVTYTNRVAD ARLGTFSQLL LQWKGSIYKL LYSEFLIFIS LYFAISLVYR LILSESQRLM FEKLALYCNS YAELIPVSFV LGFYVSLVV SRWWAQYESI PWPDRIMNLV SCNVDGEDEY GRLLRRTLMR YSNLCSVLIL RSVSTAVYKR FPSMEHVVRA G LMTPEEHK KFESLNSPHN KFWIPCVWFS NLAVKARNEG RIRDSVLLQG ILNELNTLRS QCGRLYGYDW ISIPLVYTQV VT VAVYSFF LACLIGRQFL DPEKAYPGHE LDLFVPVFTF LQFFFYAGFL KVAEQLINPF GEDDDDFETN WLIDRNLQVS LMA VDEMHQ DLPILEKDLY WNEPDPQEGE EF |
-Macromolecule #2: CALCIUM ION
Macromolecule | Name: CALCIUM ION / type: ligand / ID: 2 / Number of copies: 5 / Formula: CA |
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Molecular weight | Theoretical: 40.078 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 5 mg/mL |
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Buffer | pH: 7.5 |
Grid | Details: unspecified |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV |
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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) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Average exposure time: 10.0 sec. / Average electron dose: 76.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |