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Yorodumi- EMDB-13748: Preliminary Single particle cryo-EM reconstruction of a 40-mer as... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13748 | |||||||||
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Title | Preliminary Single particle cryo-EM reconstruction of a 40-mer assembly of Hsp26 purified from yeast. | |||||||||
Map data | Preliminary WT map of Hsp26 purified from yeast | |||||||||
Sample |
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Function / homology | Function and homology information cytoplasmic stress granule / unfolded protein binding / protein folding / cellular response to heat / mRNA binding / mitochondrion / identical protein binding / nucleus / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 10.0 Å | |||||||||
Authors | Peters C / Weinkauf S / Buchner J | |||||||||
Funding support | Germany, 1 items
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Citation | Journal: Nat Commun / Year: 2021 Title: Phosphorylation activates the yeast small heat shock protein Hsp26 by weakening domain contacts in the oligomer ensemble. Authors: Moritz Mühlhofer / Carsten Peters / Thomas Kriehuber / Marina Kreuzeder / Pamina Kazman / Natalia Rodina / Bernd Reif / Martin Haslbeck / Sevil Weinkauf / Johannes Buchner / Abstract: Hsp26 is a small heat shock protein (sHsp) from S. cerevisiae. Its chaperone activity is activated by oligomer dissociation at heat shock temperatures. Hsp26 contains 9 phosphorylation sites in ...Hsp26 is a small heat shock protein (sHsp) from S. cerevisiae. Its chaperone activity is activated by oligomer dissociation at heat shock temperatures. Hsp26 contains 9 phosphorylation sites in different structural elements. Our analysis of phospho-mimetic mutations shows that phosphorylation activates Hsp26 at permissive temperatures. The cryo-EM structure of the Hsp26 40mer revealed contacts between the conserved core domain of Hsp26 and the so-called thermosensor domain in the N-terminal part of the protein, which are targeted by phosphorylation. Furthermore, several phosphorylation sites in the C-terminal extension, which link subunits within the oligomer, are sensitive to the introduction of negative charges. In all cases, the intrinsic inhibition of chaperone activity is relieved and the N-terminal domain becomes accessible for substrate protein binding. The weakening of domain interactions within and between subunits by phosphorylation to activate the chaperone activity in response to proteotoxic stresses independent of heat stress could be a general regulation principle of sHsps. #1: Journal: Structure / Year: 2006 Title: Multiple distinct assemblies reveal conformational flexibility in the small heat shock protein Hsp26. Authors: White HE / Orlova EV / Chen S / Wang L / Ignatiou A / Gowen B / Stromer T / Franzmann TM / Haslbeck M / Buchner J / Saibil HR | |||||||||
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_13748.map.gz | 39.7 MB | EMDB map data format | |
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Header (meta data) | emd-13748-v30.xml emd-13748.xml | 13.9 KB 13.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_13748_fsc.xml | 13 KB | Display | FSC data file |
Images | emd_13748.png | 71.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13748 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13748 | HTTPS FTP |
-Validation report
Summary document | emd_13748_validation.pdf.gz | 339.9 KB | Display | EMDB validaton report |
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Full document | emd_13748_full_validation.pdf.gz | 339.4 KB | Display | |
Data in XML | emd_13748_validation.xml.gz | 11.7 KB | Display | |
Data in CIF | emd_13748_validation.cif.gz | 15.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13748 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13748 | HTTPS FTP |
-Related 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_13748.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Preliminary WT map of Hsp26 purified from yeast | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.33 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 : 40mer complex of WT Hsp26
Entire | Name: 40mer complex of WT Hsp26 |
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Components |
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-Supramolecule #1: 40mer complex of WT Hsp26
Supramolecule | Name: 40mer complex of WT Hsp26 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) / Recombinant strain: JT(DIP) GPD26(A) / Recombinant plasmid: pJV517 |
Molecular weight | Theoretical: 955.2 KDa |
-Macromolecule #1: Hsp26
Macromolecule | Name: Hsp26 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Sequence | String: MSFNSPFFDF FDNINNEVDA FNRLLGEGGL RGYAPRRQLA NTPAKDSTGK EVARPNNYAG ALYDPRDET LDDWFDNDLS LFPSGFGFPR SVAVPVDILD HDNNYELKVV VPGVKSKKDI D IEYHQNKN QILVSGEIPS TLNEESKDKV KVKESSSGKF KRVITLPDYP ...String: MSFNSPFFDF FDNINNEVDA FNRLLGEGGL RGYAPRRQLA NTPAKDSTGK EVARPNNYAG ALYDPRDET LDDWFDNDLS LFPSGFGFPR SVAVPVDILD HDNNYELKVV VPGVKSKKDI D IEYHQNKN QILVSGEIPS TLNEESKDKV KVKESSSGKF KRVITLPDYP GVDADNIKAD YA NGVLTLT VPKLKPQKDG KNHVKKIEVS SQESWGN |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R2/1 / Material: COPPER / Mesh: 400 |
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 294 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 1170 / Average exposure time: 8.0 sec. / Average electron dose: 140.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |