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Yorodumi- PDB-7pe3: Pseudo-atomic model of the tetrahedral 24mer of Hsp17 from Caenor... -
+Open data
-Basic information
Entry | Database: PDB / ID: 7pe3 | ||||||
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Title | Pseudo-atomic model of the tetrahedral 24mer of Hsp17 from Caenorhabditis elegans | ||||||
Components | SHSP domain-containing protein | ||||||
Keywords | CHAPERONE / small heat shock protein / Caenorhabditis elegans / proteostasis | ||||||
Function / homology | Alpha crystallin/Small heat shock protein, animal type / Hsp20/alpha crystallin family / Small heat shock protein (sHSP) domain profile. / Alpha crystallin/Hsp20 domain / HSP20-like chaperone / unfolded protein binding / protein refolding / response to heat / SHSP domain-containing protein Function and homology information | ||||||
Biological species | Caenorhabditis elegans (invertebrata) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 6.49 Å | ||||||
Authors | Rossa, B. / Weinkauf, S. / Buchner, J. | ||||||
Funding support | Germany, 1items
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Citation | Journal: J Biol Chem / Year: 2023 Title: The permanently chaperone-active small heat shock protein Hsp17 from Caenorhabditis elegans exhibits topological separation of its N-terminal regions. Authors: Annika Strauch / Benjamin Rossa / Fabian Köhler / Simon Haeussler / Moritz Mühlhofer / Florian Rührnößl / Caroline Körösy / Yevheniia Bushman / Barbara Conradt / Martin Haslbeck / ...Authors: Annika Strauch / Benjamin Rossa / Fabian Köhler / Simon Haeussler / Moritz Mühlhofer / Florian Rührnößl / Caroline Körösy / Yevheniia Bushman / Barbara Conradt / Martin Haslbeck / Sevil Weinkauf / Johannes Buchner / Abstract: Small Heat shock proteins (sHsps) are a family of molecular chaperones that bind nonnative proteins in an ATP-independent manner. Caenorhabditis elegans encodes 16 different sHsps, among them Hsp17, ...Small Heat shock proteins (sHsps) are a family of molecular chaperones that bind nonnative proteins in an ATP-independent manner. Caenorhabditis elegans encodes 16 different sHsps, among them Hsp17, which is evolutionarily distinct from other sHsps in the nematode. The structure and mechanism of Hsp17 and how these may differ from other sHsps remain unclear. Here, we find that Hsp17 has a distinct expression pattern, structural organization, and chaperone function. Consistent with its presence under nonstress conditions, and in contrast to many other sHsps, we determined that Hsp17 is a mono-disperse, permanently active chaperone in vitro, which interacts with hundreds of different C. elegans proteins under physiological conditions. Additionally, our cryo-EM structure of Hsp17 reveals that in the 24-mer complex, 12 N-terminal regions are involved in its chaperone function. These flexible regions are located on the outside of the spherical oligomer, whereas the other 12 N-terminal regions are engaged in stabilizing interactions in its interior. This allows the same region in Hsp17 to perform different functions depending on the topological context. Taken together, our results reveal structural and functional features that further define the structural basis of permanently active sHsps. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7pe3.cif.gz | 63.7 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7pe3.ent.gz | 45.1 KB | Display | PDB format |
PDBx/mmJSON format | 7pe3.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7pe3_validation.pdf.gz | 1.1 MB | Display | wwPDB validaton report |
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Full document | 7pe3_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | 7pe3_validation.xml.gz | 28 KB | Display | |
Data in CIF | 7pe3_validation.cif.gz | 37.7 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/pe/7pe3 ftp://data.pdbj.org/pub/pdb/validation_reports/pe/7pe3 | HTTPS FTP |
-Related structure data
Related structure data | 13346MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 17442.510 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Caenorhabditis elegans (invertebrata) / Gene: hsp-17, CELE_F52E1.7, F52E1.7 / Plasmid: pET21a / Production host: Escherichia coli BL21 (bacteria) / References: UniProt: Q7JP52 |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Tetrahedral oligomer (24mer) of Hsp17 / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Value: 0.422 MDa / Experimental value: NO |
Source (natural) | Organism: Caenorhabditis elegans (invertebrata) |
Source (recombinant) | Organism: Escherichia coli BL21 (bacteria) / Plasmid: pET21a |
Buffer solution | pH: 7.4 / Details: PBS |
Specimen | Conc.: 2.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: This sample was monodisperse |
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R2/1 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 70 % / Chamber temperature: 298 K Details: waiting time 0, blotting force 5, blotting time 2.5 |
-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 / Nominal defocus max: 2500 nm / Nominal defocus min: 500 nm / Cs: 2.7 mm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 55 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 2568 Details: Images were recorded in CDS mode as tif-stacks with 30 frames. |
Image scans | Width: 5760 / Height: 4092 |
-Processing
Software | Name: PHENIX / Version: 1.19.2_4158: / Classification: refinement | ||||||||||||||||||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 663141 Details: Gautomatch automatic picking yielded an initial dataset that was used to generate templates for cryoSPARC2 template picking. | ||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: T (tetrahedral) | ||||||||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 6.49 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 187116 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: FLEXIBLE FIT / Space: REAL | ||||||||||||||||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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