+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10618 | |||||||||
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Title | The human core BBSome complex (BBS 1,4,5,8,9,18) | |||||||||
Map data | ||||||||||
Sample |
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Keywords | ciliary transport / Arl6 effector / adaptor protein / complex / PROTEIN TRANSPORT | |||||||||
Function / homology | Function and homology information RNA polymerase II-specific DNA-binding transcription factor binding => GO:0061629 / BBSome / response to stimulus / melanosome transport / motile cilium assembly / BBSome-mediated cargo-targeting to cilium / phosphatidylinositol-3-phosphate binding / ciliary membrane / heart looping / axoneme ...RNA polymerase II-specific DNA-binding transcription factor binding => GO:0061629 / BBSome / response to stimulus / melanosome transport / motile cilium assembly / BBSome-mediated cargo-targeting to cilium / phosphatidylinositol-3-phosphate binding / ciliary membrane / heart looping / axoneme / centriolar satellite / cilium assembly / intracellular transport / visual perception / ciliary basal body / protein transport / cytosol Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.3 Å | |||||||||
Authors | Klink BU / Raunser S | |||||||||
Funding support | Germany, 1 items
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Citation | Journal: Elife / Year: 2020 Title: Structure of the human BBSome core complex. Authors: Björn Udo Klink / Christos Gatsogiannis / Oliver Hofnagel / Alfred Wittinghofer / Stefan Raunser / Abstract: The BBSome is a heterooctameric protein complex that plays a central role in primary cilia homeostasis. Its malfunction causes the severe ciliopathy Bardet-Biedl syndrome (BBS). The complex acts as a ...The BBSome is a heterooctameric protein complex that plays a central role in primary cilia homeostasis. Its malfunction causes the severe ciliopathy Bardet-Biedl syndrome (BBS). The complex acts as a cargo adapter that recognizes signaling proteins such as GPCRs and links them to the intraflagellar transport machinery. The underlying mechanism is poorly understood. Here we present a high-resolution cryo-EM structure of a human heterohexameric core subcomplex of the BBSome. The structure reveals the architecture of the complex in atomic detail. It explains how the subunits interact with each other and how disease-causing mutations hamper this interaction. The complex adopts a conformation that is open for binding to membrane-associated GTPase Arl6 and a large positively charged patch likely strengthens the interaction with the membrane. A prominent negatively charged cleft at the center of the complex is likely involved in binding of positively charged signaling sequences of cargo proteins. | |||||||||
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_10618.map.gz | 4.3 MB | EMDB map data format | |
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Header (meta data) | emd-10618-v30.xml emd-10618.xml | 13.4 KB 13.4 KB | Display Display | EMDB header |
Images | emd_10618.png | 129.9 KB | ||
Filedesc metadata | emd-10618.cif.gz | 6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10618 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10618 | HTTPS FTP |
-Validation report
Summary document | emd_10618_validation.pdf.gz | 212.8 KB | Display | EMDB validaton report |
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Full document | emd_10618_full_validation.pdf.gz | 211.9 KB | Display | |
Data in XML | emd_10618_validation.xml.gz | 6.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10618 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10618 | HTTPS FTP |
-Related structure data
Related structure data | 6xtbMC 6xt9C C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_10618.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Human BBSome core complex
Entire | Name: Human BBSome core complex |
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Components |
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-Supramolecule #1: Human BBSome core complex
Supramolecule | Name: Human BBSome core complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: BBSome core complex containing BBS1,4,5,8,9 and 18. Only BBS5 was modelled into this map since we obtained another map with higher resolution for the other subunits (see related entry) |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Bardet-Biedl syndrome 5 protein
Macromolecule | Name: Bardet-Biedl syndrome 5 protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 38.797926 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MSVLDALWED RDVRFDLSAQ QMKTRPGEVL IDCLDSIEDT KGNNGDRGRL LVTNLRILWH SLALSRVNVS VGYNCILNIT TRTANSKLR GQTEALYILT KCNSTRFEFI FTNLVPGSPR LFTSVMAVHR AYETSKMYRD FKLRSALIQN KQLRLLPQEH V YDKINGVW ...String: MSVLDALWED RDVRFDLSAQ QMKTRPGEVL IDCLDSIEDT KGNNGDRGRL LVTNLRILWH SLALSRVNVS VGYNCILNIT TRTANSKLR GQTEALYILT KCNSTRFEFI FTNLVPGSPR LFTSVMAVHR AYETSKMYRD FKLRSALIQN KQLRLLPQEH V YDKINGVW NLSSDQGNLG TFFITNVRIV WHANMNDSFN VSIPYLQIRS IKIRDSKFGL ALVIESSQQS GGYVLGFKID PV EKLQESV KEINSLHKVY SASPIFGVDY EMEEKPQPLE ALTVEQIQDD VEIDSDGHTD AFVAYFADGN KQQDREPVFS EEL GLAIEK LKDGFTLQGL WEVMS UniProtKB: Bardet-Biedl syndrome 5 protein |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.08 mg/mL | ||||||||
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Buffer | pH: 7.5 Component:
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 286 K / Instrument: FEI VITROBOT MARK III Details: double blot with 2 minutes incubation after first sample application. | ||||||||
Details | The sample was cross linked with 0.5% glutaraldehyde |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-50 / Number real images: 15266 / Average exposure time: 15.0 sec. / Average electron dose: 67.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated defocus max: 1.0 µm / Calibrated defocus min: 0.3 µm / Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |