+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21144 | |||||||||
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Title | Structure of the bovine BBSome complex | |||||||||
Map data | Post-processed map of the bovine BBSome | |||||||||
Sample |
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Keywords | Cilia / ciliopathy / complex / membrane-protein transport / PROTEIN TRANSPORT | |||||||||
Function / homology | Function and homology information establishment of anatomical structure orientation / BBSome-mediated cargo-targeting to cilium / multi-ciliated epithelial cell differentiation / receptor localization to non-motile cilium / renal tubule development / BBSome / camera-type eye photoreceptor cell differentiation / smoothened binding / establishment of planar polarity / photoreceptor connecting cilium ...establishment of anatomical structure orientation / BBSome-mediated cargo-targeting to cilium / multi-ciliated epithelial cell differentiation / receptor localization to non-motile cilium / renal tubule development / BBSome / camera-type eye photoreceptor cell differentiation / smoothened binding / establishment of planar polarity / photoreceptor connecting cilium / inner ear receptor cell stereocilium organization / patched binding / olfactory bulb development / protein localization to cilium / non-motile cilium assembly / phosphatidylinositol-3-phosphate binding / establishment of epithelial cell apical/basal polarity / non-motile cilium / regulation of stress fiber assembly / motile cilium / centrosome cycle / ciliary membrane / pericentriolar material / fat cell differentiation / axoneme / intracellular transport / cilium assembly / centriolar satellite / ciliary basal body / axon guidance / protein localization to plasma membrane / multicellular organism growth / protein localization / cilium / regulation of protein localization / sensory perception of smell / protein-macromolecule adaptor activity / protein transport / RNA polymerase II-specific DNA-binding transcription factor binding / neuron projection / centrosome / membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | Bos taurus (cattle) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
Authors | Singh SK / Gui M | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Elife / Year: 2020 Title: Structure and activation mechanism of the BBSome membrane protein trafficking complex. Authors: Sandeep K Singh / Miao Gui / Fujiet Koh / Matthew Cj Yip / Alan Brown / Abstract: Bardet-Biedl syndrome (BBS) is a currently incurable ciliopathy caused by the failure to correctly establish or maintain cilia-dependent signaling pathways. Eight proteins associated with BBS ...Bardet-Biedl syndrome (BBS) is a currently incurable ciliopathy caused by the failure to correctly establish or maintain cilia-dependent signaling pathways. Eight proteins associated with BBS assemble into the BBSome, a key regulator of the ciliary membrane proteome. We report the electron cryomicroscopy (cryo-EM) structures of the native bovine BBSome in inactive and active states at 3.1 and 3.5 Å resolution, respectively. In the active state, the BBSome is bound to an Arf-family GTPase (ARL6/BBS3) that recruits the BBSome to ciliary membranes. ARL6 recognizes a composite binding site formed by BBS1 and BBS7 that is occluded in the inactive state. Activation requires an unexpected swiveling of the β-propeller domain of BBS1, the subunit most frequently implicated in substrate recognition, which widens a central cavity of the BBSome. Structural mapping of disease-causing mutations suggests that pathogenesis results from folding defects and the disruption of autoinhibition and activation. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21144.map.gz | 9.1 MB | EMDB map data format | |
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Header (meta data) | emd-21144-v30.xml emd-21144.xml | 37.8 KB 37.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_21144_fsc.xml | 11.4 KB | Display | FSC data file |
Images | emd_21144.png | 197.7 KB | ||
Masks | emd_21144_msk_1.map emd_21144_msk_2.map emd_21144_msk_3.map | 125 MB 125 MB 125 MB | Mask map | |
Filedesc metadata | emd-21144.cif.gz | 9.5 KB | ||
Others | emd_21144_additional_1.map.gz emd_21144_additional_2.map.gz emd_21144_additional_3.map.gz emd_21144_additional_4.map.gz emd_21144_half_map_1.map.gz emd_21144_half_map_2.map.gz | 3.4 MB 8.2 MB 9.1 MB 98.6 MB 98.9 MB 98.9 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21144 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21144 | HTTPS FTP |
-Related structure data
Related structure data | 6vbuMC 6vbvC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21144.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Post-processed map of the bovine BBSome | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_21144_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Mask #2
File | emd_21144_msk_2.map | ||||||||||||
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Density Histograms |
-Mask #3
File | emd_21144_msk_3.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Composite post-processed map following multibody refinement
File | emd_21144_additional_1.map | ||||||||||||
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Annotation | Composite post-processed map following multibody refinement | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Post-processed map of the BBSome body following multibody refinement
File | emd_21144_additional_2.map | ||||||||||||
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Annotation | Post-processed map of the BBSome body following multibody refinement | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Post-processed map of the BBSome
File | emd_21144_additional_3.map | ||||||||||||
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Annotation | Post-processed map of the BBSome | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unfiltered map of the BBSome
File | emd_21144_additional_4.map | ||||||||||||
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Annotation | Unfiltered map of the BBSome | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1
File | emd_21144_half_map_1.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2
File | emd_21144_half_map_2.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Bovine BBSome complex
+Supramolecule #1: Bovine BBSome complex
+Macromolecule #1: Bardet-Biedl syndrome 18 protein
+Macromolecule #2: BBS1 domain-containing protein
+Macromolecule #3: Bardet-Biedl syndrome 2 protein homolog
+Macromolecule #4: Bardet-Biedl syndrome 4 protein homolog
+Macromolecule #5: Bardet-Biedl syndrome 5 protein homolog
+Macromolecule #6: Bardet-Biedl syndrome 7 protein homolog
+Macromolecule #7: Bardet-Biedl syndrome 8 protein
+Macromolecule #8: Bardet-Biedl syndrome 9
+Macromolecule #9: CALCIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.7 mg/mL | |||||||||||||||
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Buffer | pH: 7.5 Component:
Details: The buffer also contained 36 uM ARL6 and 1 mM GTP. | |||||||||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR Details: The grids were glow discharged at 15 mA for 30 s with a PELCO easiGlow Glow Discharge Cleaning System. | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK II / Details: Grids were blotted for 2 s with a -2 offset.. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.4 µm / Nominal defocus min: 1.1 µm / Nominal magnification: 81000 |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 25 eV |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 2 / Number real images: 9408 / Average exposure time: 4.0 sec. / Average electron dose: 56.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
-Atomic model buiding 1
Details | During refinement, the resolution limit was set to 3.1 Angstrom. Secondary structure, Ramachandran and rotamer restraints were applied during refinement. |
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Refinement | Space: REAL / Protocol: OTHER / Overall B value: 43.6 / Target criteria: Correlation coefficient |
Output model | PDB-6vbu: |