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Yorodumi- EMDB-37863: Cryo-EM structure of H. thermophilus GroEL-GroES bullet complex -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-37863 | |||||||||
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Title | Cryo-EM structure of H. thermophilus GroEL-GroES bullet complex | |||||||||
Map data | ||||||||||
Sample |
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Keywords | Chaperone / Chaperonin / ATPase / protein folding | |||||||||
Function / homology | Function and homology information chaperonin ATPase / protein folding chaperone / isomerase activity / ATP-dependent protein folding chaperone / unfolded protein binding / protein refolding / ATP binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Hydrogenobacter thermophilus TK-6 (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.6 Å | |||||||||
Authors | Liao Z / Gopalasingam CC / Kameya M / Gerle C / Shigematsu H / Ishii M / Arakawa T / Fushinobu S | |||||||||
Funding support | Japan, 2 items
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Citation | Journal: Structure / Year: 2024 Title: Structural insights into thermophilic chaperonin complexes. Authors: Zengwei Liao / Chai C Gopalasingam / Masafumi Kameya / Christoph Gerle / Hideki Shigematsu / Masaharu Ishii / Takatoshi Arakawa / Shinya Fushinobu / Abstract: Group I chaperonins are dual heptamer protein complexes that play significant roles in protein homeostasis. The structure and function of the Escherichia coli chaperonin are well characterized. ...Group I chaperonins are dual heptamer protein complexes that play significant roles in protein homeostasis. The structure and function of the Escherichia coli chaperonin are well characterized. However, the dynamic properties of chaperonins, such as large ATPase-dependent conformational changes by binding of lid-like co-chaperonin GroES, have made structural analyses challenging, and our understanding of these changes during the turnover of chaperonin complex formation is limited. In this study, we used single-particle cryogenic electron microscopy to investigate the structures of GroES-bound chaperonin complexes from the thermophilic hydrogen-oxidizing bacteria Hydrogenophilus thermoluteolus and Hydrogenobacter thermophilus in the presence of ATP and AMP-PNP. We captured the structure of an intermediate state chaperonin complex, designated as an asymmetric football-shaped complex, and performed analyses to decipher the dynamic structural variations. Our structural analyses of inter- and intra-subunit communications revealed a unique mechanism of complex formation through the binding of a second GroES to a bullet-shaped complex. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_37863.map.gz | 256.4 MB | EMDB map data format | |
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Header (meta data) | emd-37863-v30.xml emd-37863.xml | 26 KB 26 KB | Display Display | EMDB header |
Images | emd_37863.png | 86.1 KB | ||
Masks | emd_37863_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-37863.cif.gz | 6.8 KB | ||
Others | emd_37863_additional_1.map.gz emd_37863_additional_2.map.gz emd_37863_half_map_1.map.gz emd_37863_half_map_2.map.gz | 484.1 MB 456.8 MB 474.4 MB 474.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-37863 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-37863 | HTTPS FTP |
-Validation report
Summary document | emd_37863_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_37863_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_37863_validation.xml.gz | 19 KB | Display | |
Data in CIF | emd_37863_validation.cif.gz | 22.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-37863 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-37863 | HTTPS FTP |
-Related structure data
Related structure data | 8wuxMC 8wu4C 8wucC 8wuwC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_37863.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.752 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_37863_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: #1
File | emd_37863_additional_1.map | ||||||||||||
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Density Histograms |
-Additional map: #2
File | emd_37863_additional_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_37863_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_37863_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : H. thermophilus GroEL-GroES bullet complex
Entire | Name: H. thermophilus GroEL-GroES bullet complex |
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Components |
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-Supramolecule #1: H. thermophilus GroEL-GroES bullet complex
Supramolecule | Name: H. thermophilus GroEL-GroES bullet complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: Hydrogenobacter thermophilus TK-6 (bacteria) |
-Supramolecule #2: chaperonin GroEL
Supramolecule | Name: chaperonin GroEL / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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-Supramolecule #3: co-chaperonin GroES
Supramolecule | Name: co-chaperonin GroES / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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-Macromolecule #1: Chaperonin GroEL
Macromolecule | Name: Chaperonin GroEL / type: protein_or_peptide / ID: 1 / Number of copies: 14 / Enantiomer: LEVO / EC number: chaperonin ATPase |
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Source (natural) | Organism: Hydrogenobacter thermophilus TK-6 (bacteria) / Strain: TK-6 |
Molecular weight | Theoretical: 56.600918 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: AAKKVIYGED ARARLKAGVD KLANAVKVTL GPRGREVIIE KKWGTPVVTK DGVTVAKEIE FKDPYENMGA QLVKEVASKT SDVAGDGTT TATVLAQAIF NEGLRAIASG ANPMDIKRGI DKAVETVVNE IKKLSIPVSG RKEIEQVATI SANNDATIGK I IADAMEAV ...String: AAKKVIYGED ARARLKAGVD KLANAVKVTL GPRGREVIIE KKWGTPVVTK DGVTVAKEIE FKDPYENMGA QLVKEVASKT SDVAGDGTT TATVLAQAIF NEGLRAIASG ANPMDIKRGI DKAVETVVNE IKKLSIPVSG RKEIEQVATI SANNDATIGK I IADAMEAV GKDGVITVEE SKSAETTLET VQGMQFDRGY LSPYFVTNPD KMEAVLEDPF ILIYEKKISN VKDLLPVLEN VV RAGKPLL IIAEDVEAEA LATLVVNHIK GVIRACAVKA PGFGQRRKDY LQDIAILTGG TAITEELGIK LESVTLDMLG RAD KVIVDK DNTTIVGGKG SKEAIQARIE QIKRQILETT SDYDREKLQE RLAKLSGGVA IIRVGAATEA ELKEKKARVE DAVH ATKAA VEEGIVPGGG VALVRASEAL DNLKVDNADQ QLGIDIIKKA CRTPIRQIAA NSGFEGYVVL EKVLQLGKEK GKNWG FDAG VGDYKDMVEA GIIDPTKVVR VAIQNAASVA GTMLTAEALV AEIPE UniProtKB: Chaperonin GroEL |
-Macromolecule #2: Co-chaperonin GroES
Macromolecule | Name: Co-chaperonin GroES / type: protein_or_peptide / ID: 2 / Number of copies: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: Hydrogenobacter thermophilus TK-6 (bacteria) / Strain: TK-6 |
Molecular weight | Theoretical: 10.440112 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: RLRPLYDKIV VKRMEEQEQK TPSGIIIPDT AKEKPQIGEV IAVGDGKLLS NGQIVSPKVK KGDKVVFNKY AGTEVELDGE KYLIMSEDE VLAVI UniProtKB: Co-chaperonin GroES |
-Macromolecule #3: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 3 / Number of copies: 14 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Macromolecule #4: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
Macromolecule | Name: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / type: ligand / ID: 4 / Number of copies: 14 / Formula: ANP |
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Molecular weight | Theoretical: 506.196 Da |
Chemical component information | ChemComp-ANP: |
-Macromolecule #5: POTASSIUM ION
Macromolecule | Name: POTASSIUM ION / type: ligand / ID: 5 / Number of copies: 7 / Formula: K |
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Molecular weight | Theoretical: 39.098 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 10 sec. / Pretreatment - Atmosphere: OTHER / Pretreatment - Pressure: 0.007 kPa Details: glow discharged at 7 Pa, 10 mA, 10 sec by a JEOL JEC-3000FC auto fine coater | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 281 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | JEOL CRYO ARM 300 |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 5650 / Average exposure time: 2.264 sec. / Average electron dose: 49.97 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.6 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 60000 |
Sample stage | Cooling holder cryogen: NITROGEN |