+Open data
-Basic information
Entry | Database: PDB / ID: 7s8x | ||||||
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Title | Cryo-EM Structure of dolphin Prestin: Sensor Up (compact) state | ||||||
Components | Prestin | ||||||
Keywords | MOTOR PROTEIN / Outer hair cells / electromotility / mechanotransduction / hearing / deafness / frequency sensation / echolocation / SLC26 / SLC26A5 | ||||||
Function / homology | Function and homology information cochlear outer hair cell electromotile response / oxalate transmembrane transporter activity / secondary active sulfate transmembrane transporter activity / bicarbonate transmembrane transporter activity / chloride transmembrane transporter activity / sensory perception of sound / regulation of cell shape / plasma membrane Similarity search - Function | ||||||
Biological species | Tursiops truncatus (common bottlenose dolphin) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.3 Å | ||||||
Authors | Bavi, N. / Clark, M.D. / Contreras, G.F. / Shen, R. / Reddy, B.G. / Milewski, W. / Perozo, E. | ||||||
Funding support | United States, 1items
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Citation | Journal: Nature / Year: 2021 Title: The conformational cycle of prestin underlies outer-hair cell electromotility. Authors: Navid Bavi / Michael David Clark / Gustavo F Contreras / Rong Shen / Bharat G Reddy / Wieslawa Milewski / Eduardo Perozo / Abstract: The voltage-dependent motor protein prestin (also known as SLC26A5) is responsible for the electromotive behaviour of outer-hair cells and underlies the cochlear amplifier. Knockout or impairment of ...The voltage-dependent motor protein prestin (also known as SLC26A5) is responsible for the electromotive behaviour of outer-hair cells and underlies the cochlear amplifier. Knockout or impairment of prestin causes severe hearing loss. Despite the key role of prestin in hearing, the mechanism by which mammalian prestin senses voltage and transduces it into cellular-scale movements (electromotility) is poorly understood. Here we determined the structure of dolphin prestin in six distinct states using single-particle cryo-electron microscopy. Our structural and functional data suggest that prestin adopts a unique and complex set of states, tunable by the identity of bound anions (Cl or SO). Salicylate, a drug that can cause reversible hearing loss, competes for the anion-binding site of prestin, and inhibits its function by immobilizing prestin in a new conformation. Our data suggest that the bound anion together with its coordinating charged residues and helical dipole act as a dynamic voltage sensor. An analysis of all of the anion-dependent conformations reveals how structural rearrangements in the voltage sensor are coupled to conformational transitions at the protein-membrane interface, suggesting a previously undescribed mechanism of area expansion. Visualization of the electromotility cycle of prestin distinguishes the protein from the closely related SLC26 anion transporters, highlighting the basis for evolutionary specialization of the mammalian cochlear amplifier at a high resolution. | ||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7s8x.cif.gz | 233.1 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7s8x.ent.gz | 189.3 KB | Display | PDB format |
PDBx/mmJSON format | 7s8x.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7s8x_validation.pdf.gz | 928.3 KB | Display | wwPDB validaton report |
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Full document | 7s8x_full_validation.pdf.gz | 945.6 KB | Display | |
Data in XML | 7s8x_validation.xml.gz | 44.5 KB | Display | |
Data in CIF | 7s8x_validation.cif.gz | 68 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/s8/7s8x ftp://data.pdbj.org/pub/pdb/validation_reports/s8/7s8x | HTTPS FTP |
-Related structure data
Related structure data | 24928MC 7s9aC 7s9bC 7s9cC 7s9dC 7s9eC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 80973.750 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Tursiops truncatus (common bottlenose dolphin) Gene: SLC26A5 / Cell line (production host): Hek 293 / Production host: Homo sapiens (human) / References: UniProt: D7PC76 |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: CELL / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Cryo-EM Structure of dolphin Prestin: Sensor Up (compact) state Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: Tursiops truncatus (common bottlenose dolphin) |
Source (recombinant) | Organism: Homo sapiens (human) / Cell: Hek 293 |
Buffer solution | pH: 7.4 Details: 360 mM NaCl, 20 mM Tris-Hcl, 3mM DTT, 1mM EDTA, 0.02 % GDN |
Buffer component | Conc.: 360 mM / Name: sodium chloride / Formula: NaCl |
Specimen | Conc.: 2.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: Monodisperse peak at around 14 ml using SEC (Superose 6 column) |
Specimen support | Details: Grids were plasma cleaned for 30 s in an air mixture in a Solarus Plasma Cleaner (Gatan). Grid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 295 K Details: 4.5 s blot times, blot force 3, and double filter papers on each side of the vitrobot. |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 1 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
CTF correction | Type: NONE |
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3D reconstruction | Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 206000 / Symmetry type: POINT |