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- EMDB-23002: Cryo-EM structure of the CRAC channel Orai in an open conformatio... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-23002 | |||||||||
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Title | Cryo-EM structure of the CRAC channel Orai in an open conformation; H206A gain-of-function mutation in complex with an antibody | |||||||||
![]() | cryo-EM map | |||||||||
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Function / homology | ![]() store-operated calcium entry / store-operated calcium channel activity / positive regulation of calcineurin-NFAT signaling cascade / positive regulation of calcium ion transport / calcium channel regulator activity / calcium channel complex / calcium ion transmembrane transport / calcium channel activity / nervous system development / identical protein binding ...store-operated calcium entry / store-operated calcium channel activity / positive regulation of calcineurin-NFAT signaling cascade / positive regulation of calcium ion transport / calcium channel regulator activity / calcium channel complex / calcium ion transmembrane transport / calcium channel activity / nervous system development / identical protein binding / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
![]() | Long SB / Hou X / Outhwaite IR | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of the calcium release-activated calcium channel Orai in an open conformation. Authors: Xiaowei Hou / Ian R Outhwaite / Leanne Pedi / Stephen Barstow Long / ![]() Abstract: The calcium release-activated calcium channel Orai regulates Ca entry into non-excitable cells and is required for proper immune function. While the channel typically opens following Ca release from ...The calcium release-activated calcium channel Orai regulates Ca entry into non-excitable cells and is required for proper immune function. While the channel typically opens following Ca release from the endoplasmic reticulum, certain pathologic mutations render the channel constitutively open. Previously, using one such mutation (H206A), we obtained low (6.7 Å) resolution X-ray structural information on Orai in an open conformation (Hou et al., 2018). Here we present a structure of this open conformation at 3.3 Å resolution using fiducial-assisted cryo-electron microscopy. The improved structure reveals the conformations of amino acids in the open pore, which dilates by outward movements of subunits. A ring of phenylalanine residues repositions to expose previously shielded glycine residues to the pore without significant rotational movement of the associated helices. Together with other hydrophobic amino acids, the phenylalanines act as the channel's gate. Structured M1-M2 turrets, not evident previously, form the channel's extracellular entrance. | |||||||||
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 | ![]() | 67.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 15.3 KB 15.3 KB | Display Display | ![]() |
Images | ![]() | 199.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 456.5 KB | Display | ![]() |
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Full document | ![]() | 456 KB | Display | |
Data in XML | ![]() | 6.4 KB | Display | |
Data in CIF | ![]() | 7.3 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7kr5MC 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
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | cryo-EM map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.87078 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 : Orai channel, H206A mutant, in complex with three Fab fragments
Entire | Name: Orai channel, H206A mutant, in complex with three Fab fragments |
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Components |
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-Supramolecule #1: Orai channel, H206A mutant, in complex with three Fab fragments
Supramolecule | Name: Orai channel, H206A mutant, in complex with three Fab fragments type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 |
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-Supramolecule #2: Orai channel, H206A mutant
Supramolecule | Name: Orai channel, H206A mutant / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() |
-Supramolecule #3: Fab fragments
Supramolecule | Name: Fab fragments / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Macromolecule #1: Calcium release-activated calcium channel protein 1
Macromolecule | Name: Calcium release-activated calcium channel protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 24.160168 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MSQSGEDLHS PTYLSWRKLQ LSRAKLKASS KTSALLSGFA MVAMVEVQLD HDTNVPPGML IAFAICTTLL VAVAMLALMI STCILPNIE TVSNLHSISL VHESPHERLH WYIETAWAFS TLLGLILFLL EIAILCWVKF YDLSPPAAWS ATVVLIPVMI I FMAFAIHF ...String: MSQSGEDLHS PTYLSWRKLQ LSRAKLKASS KTSALLSGFA MVAMVEVQLD HDTNVPPGML IAFAICTTLL VAVAMLALMI STCILPNIE TVSNLHSISL VHESPHERLH WYIETAWAFS TLLGLILFLL EIAILCWVKF YDLSPPAAWS ATVVLIPVMI I FMAFAIHF YRSLVSHKYE VTVSGIRELE MLKEQMEQDH LEHHNNIRNN GEGEEF |
-Macromolecule #2: 19B5 Fab heavy chain
Macromolecule | Name: 19B5 Fab heavy chain / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 26.886477 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTLNMLLGLK WVFFVVFYQG VHCEVQLVES GGGLVQPKGS LKLSCAASGF TFNTYAMHWV RQAPGKGLEW VARIRTKSNN YATYYADSV KDRFTISRDD SQSMLYLQMN NLKTEDTAMY YCVRQKYGNY FDYWGQGTTL TVSSAKTTPP SVYPLAPGSA A QTNSMVTL ...String: MTLNMLLGLK WVFFVVFYQG VHCEVQLVES GGGLVQPKGS LKLSCAASGF TFNTYAMHWV RQAPGKGLEW VARIRTKSNN YATYYADSV KDRFTISRDD SQSMLYLQMN NLKTEDTAMY YCVRQKYGNY FDYWGQGTTL TVSSAKTTPP SVYPLAPGSA A QTNSMVTL GCLVKGYFPE PVTVTWNSGS LSSGVHTFPA VLQSDLYTLS SSVTVPSSSW PSETVTCNVA HPASSTKVDK KI VPRD |
-Macromolecule #3: 19B5 Fab light chain
Macromolecule | Name: 19B5 Fab light chain / type: protein_or_peptide / ID: 3 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 25.587295 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MESQTQVFVY MLLWLSGVDG DVVMTQSQKF MSTSVGDRVS VTCKASQNVG INVAWYQQKP GQSPKALINS ASYRNSGVPD RFTGGGSGT DFTLTINNVQ SEDLAEYFCQ QCNSYPLTFG AGTKLELRRA DAAPTVSIFP PSSEQLTSGG ASVVCFLNNF Y PKDINVKW ...String: MESQTQVFVY MLLWLSGVDG DVVMTQSQKF MSTSVGDRVS VTCKASQNVG INVAWYQQKP GQSPKALINS ASYRNSGVPD RFTGGGSGT DFTLTINNVQ SEDLAEYFCQ QCNSYPLTFG AGTKLELRRA DAAPTVSIFP PSSEQLTSGG ASVVCFLNNF Y PKDINVKW KIDGSERQNG VLNSWTDQDS KDSTYSMSST LTLTKDEYER HNSYTCEATH KTSTSPIVKS FNRN |
-Macromolecule #4: CALCIUM ION
Macromolecule | Name: CALCIUM ION / type: ligand / ID: 4 / Number of copies: 1 / 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
Buffer | pH: 8.5 |
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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) / 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: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |