+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-25046 | |||||||||
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Title | Tomogram of mPCDH15/39G7-AuNP complex | |||||||||
Map data | Tomogram of recombinant mPCDH15 extracellular domain in complex with a AuNP/39G7-Fab' conjugate | |||||||||
Sample |
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Keywords | cadherin / antibody / hearing / mechanotransduction / CELL ADHESION | |||||||||
Biological species | Oryctolagus cuniculus (rabbit) / Mus musculus (house mouse) | |||||||||
Method | electron tomography / cryo EM | |||||||||
Authors | Elferich J / Clark S / Ge J / Goehring A / Matsui A / Gouaux E | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Elife / Year: 2021 Title: Molecular structures and conformations of protocadherin-15 and its complexes on stereocilia elucidated by cryo-electron tomography. Authors: Johannes Elferich / Sarah Clark / Jingpeng Ge / April Goehring / Aya Matsui / Eric Gouaux / Abstract: Mechanosensory transduction (MT), the conversion of mechanical stimuli into electrical signals, underpins hearing and balance and is carried out within hair cells in the inner ear. Hair cells harbor ...Mechanosensory transduction (MT), the conversion of mechanical stimuli into electrical signals, underpins hearing and balance and is carried out within hair cells in the inner ear. Hair cells harbor actin-filled stereocilia, arranged in rows of descending heights, where the tips of stereocilia are connected to their taller neighbors by a filament composed of protocadherin 15 (PCDH15) and cadherin 23 (CDH23), deemed the 'tip link.' Tension exerted on the tip link opens an ion channel at the tip of the shorter stereocilia, thus converting mechanical force into an electrical signal. While biochemical and structural studies have provided insights into the molecular composition and structure of isolated portions of the tip link, the architecture, location, and conformational states of intact tip links, on stereocilia, remains unknown. Here, we report in situ cryo-electron microscopy imaging of the tip link in mouse stereocilia. We observe individual PCDH15 molecules at the tip and shaft of stereocilia and determine their stoichiometry, conformational heterogeneity, and their complexes with other filamentous proteins, perhaps including CDH23. The PCDH15 complexes occur in clusters, frequently with more than one copy of PCDH15 at the tip of stereocilia, suggesting that tip links might consist of more than one copy of PCDH15 complexes and, by extension, might include multiple MT complexes. #1: Journal: Biorxiv / Year: 2021 Title: Molecular structure and conformation of stereocilia tip-links elucidated by cryo-electron tomography Authors: Elferich J / Clark S / Ge J / Goehring A / Matsui A / Gouaux E | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_25046.map.gz | 172.6 MB | EMDB map data format | |
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Header (meta data) | emd-25046-v30.xml emd-25046.xml | 11.9 KB 11.9 KB | Display Display | EMDB header |
Images | emd_25046.png | 151 KB | ||
Filedesc metadata | emd-25046.cif.gz | 3.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-25046 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25046 | HTTPS FTP |
-Validation report
Summary document | emd_25046_validation.pdf.gz | 405.5 KB | Display | EMDB validaton report |
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Full document | emd_25046_full_validation.pdf.gz | 405 KB | Display | |
Data in XML | emd_25046_validation.xml.gz | 4.8 KB | Display | |
Data in CIF | emd_25046_validation.cif.gz | 5.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25046 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25046 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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EM raw data | EMPIAR-10820 (Title: Molecular structure and conformation of stereocilia tip-links elucidated by cryo-electron tomography Data size: 58.5 Data #1: Tiltseries of mPCDH15/39G7-AuNP complex [tilt series] Data #2: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #3: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #4: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #5: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #6: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #7: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #8: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #9: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #10: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #11: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #12: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #13: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #14: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #15: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series] Data #16: Tiltseries of mouse stereocilia containing PCDH15 molecules labeled with 39G7-AuNPs [tilt series]) EMPIAR-10898 (Title: Cryo-electron tilt series of mouse stereocilia / Data size: 1.9 TB Data #1: Unaligned tilt series of mouse stereocilia [tilt series]) |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_25046.map.gz / Format: CCP4 / Size: 321.9 MB / Type: IMAGE STORED AS SIGNED BYTE | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Tomogram of recombinant mPCDH15 extracellular domain in complex with a AuNP/39G7-Fab' conjugate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.756 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 : mPCDH15/39G7-AuNP complex
Entire | Name: mPCDH15/39G7-AuNP complex |
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Components |
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-Supramolecule #1: mPCDH15/39G7-AuNP complex
Supramolecule | Name: mPCDH15/39G7-AuNP complex / type: complex / ID: 1 / Parent: 0 |
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-Supramolecule #2: 39G7-AuNP conjugate
Supramolecule | Name: 39G7-AuNP conjugate / type: complex / ID: 2 / Parent: 1 |
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Source (natural) | Organism: Oryctolagus cuniculus (rabbit) |
-Supramolecule #3: mPCDH15 extracellular domain
Supramolecule | Name: mPCDH15 extracellular domain / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: Mus musculus (house mouse) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
Sectioning | Other: NO SECTIONING |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number real images: 41 / Average electron dose: 3.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Algorithm: BACK PROJECTION / Software - Name: IMOD / Number images used: 41 |
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