+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-45080 | ||||||||||||
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Title | E.Faecium GroEL | ||||||||||||
Map data | |||||||||||||
Sample |
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Keywords | GroEL / Chaperone / ANTIBIOTIC | ||||||||||||
Function / homology | Function and homology information mitochondrial unfolded protein response / protein import into mitochondrial intermembrane space / GroEL-GroES complex / chaperonin ATPase / positive regulation of interferon-alpha production / chaperone cofactor-dependent protein refolding / isomerase activity / ATP-dependent protein folding chaperone / positive regulation of interleukin-6 production / positive regulation of type II interferon production ...mitochondrial unfolded protein response / protein import into mitochondrial intermembrane space / GroEL-GroES complex / chaperonin ATPase / positive regulation of interferon-alpha production / chaperone cofactor-dependent protein refolding / isomerase activity / ATP-dependent protein folding chaperone / positive regulation of interleukin-6 production / positive regulation of type II interferon production / positive regulation of T cell activation / unfolded protein binding / protein-folding chaperone binding / response to heat / protein refolding / ATP binding Similarity search - Function | ||||||||||||
Biological species | Enterococcus faecium (bacteria) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.97 Å | ||||||||||||
Authors | Watson ER / Lander GC | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: J Am Chem Soc / Year: 2024 Title: Bis-sulfonamido-2-phenylbenzoxazoles Validate the GroES/EL Chaperone System as a Viable Antibiotic Target. Authors: Jack Godek / Jared Sivinski / Edmond R Watson / Felicidad Lebario / Wenli Xu / Mckayla Stevens / Christopher J Zerio / Andrew J Ambrose / Xiaoyi Zhu / Carlee A Trindl / Donna D Zhang / ...Authors: Jack Godek / Jared Sivinski / Edmond R Watson / Felicidad Lebario / Wenli Xu / Mckayla Stevens / Christopher J Zerio / Andrew J Ambrose / Xiaoyi Zhu / Carlee A Trindl / Donna D Zhang / Steven M Johnson / Gabriel C Lander / Eli Chapman / Abstract: We recently reported on small-molecule inhibitors of the GroES/GroEL chaperone system as potential antibiotics against and the ESKAPE pathogens but were unable to establish GroES/GroEL as the ...We recently reported on small-molecule inhibitors of the GroES/GroEL chaperone system as potential antibiotics against and the ESKAPE pathogens but were unable to establish GroES/GroEL as the cellular target, leading to cell death. In this study, using two of our most potent -sulfonamido-2-phenylbenzoxazoles (PBZs), we established the binding site of the PBZ molecules using cryo-EM and found that GroEL was the cellular target responsible for the mode of action. Cryo-EM revealed that PBZ1587 binds at the GroEL ring-ring interface (RRI). A cellular reporter assay confirmed that PBZ1587 engaged GroEL in cells, but cellular rescue experiments showed potential off-target effects. This prompted us to explore a closely related analogue, PBZ1038, which is also bound to the RRI. Biochemical characterization showed potent inhibition of Gram-negative chaperonins but much lower potency of chaperonin from a Gram-positive organism, . A cellular reporter assay showed that PBZ1038 also engaged GroEL in cells and that the cytotoxic phenotype could be rescued by a chromosomal copy of GroEL/GroES or by expressing a recalcitrant RRI mutant. These data argue that PBZ1038's antimicrobial action is exerted through inhibition of GroES/GroEL, validating this chaperone system as an antibiotic target. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_45080.map.gz | 59.4 MB | EMDB map data format | |
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Header (meta data) | emd-45080-v30.xml emd-45080.xml | 16.2 KB 16.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_45080_fsc.xml | 8.5 KB | Display | FSC data file |
Images | emd_45080.png | 246.5 KB | ||
Filedesc metadata | emd-45080.cif.gz | 5.5 KB | ||
Others | emd_45080_additional_1.map.gz emd_45080_half_map_1.map.gz emd_45080_half_map_2.map.gz | 31.7 MB 59.4 MB 59.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-45080 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-45080 | HTTPS FTP |
-Validation report
Summary document | emd_45080_validation.pdf.gz | 1005.4 KB | Display | EMDB validaton report |
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Full document | emd_45080_full_validation.pdf.gz | 1005 KB | Display | |
Data in XML | emd_45080_validation.xml.gz | 16.5 KB | Display | |
Data in CIF | emd_45080_validation.cif.gz | 21.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-45080 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-45080 | HTTPS FTP |
-Related structure data
Related structure data | 9c0dMC 9c0bC 9c0cC M: atomic model generated by this map C: citing same article (ref.) |
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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_45080.map.gz / Format: CCP4 / Size: 64 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: 1.15 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: #1
File | emd_45080_additional_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_45080_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_45080_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : E.faecium GroEL
Entire | Name: E.faecium GroEL |
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Components |
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-Supramolecule #1: E.faecium GroEL
Supramolecule | Name: E.faecium GroEL / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Enterococcus faecium (bacteria) |
-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: Enterococcus faecium (bacteria) |
Molecular weight | Theoretical: 57.243074 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MAKELKFAED ARAAMLRGVD KLADTVKVTL GPKGRNVVLE KSYGSPLITN DGVTIAKEIE LEDHFENMGA KLVSEVASKT NDIAGDGTT TATVLTQAIV REGLKNVTAG ANPLGIRRGI ELATKAAVEE LHNISTVVDS KEAIAQVAAV SSGSDKVGHL I ADAMEKVG ...String: MAKELKFAED ARAAMLRGVD KLADTVKVTL GPKGRNVVLE KSYGSPLITN DGVTIAKEIE LEDHFENMGA KLVSEVASKT NDIAGDGTT TATVLTQAIV REGLKNVTAG ANPLGIRRGI ELATKAAVEE LHNISTVVDS KEAIAQVAAV SSGSDKVGHL I ADAMEKVG NDGVITIEES KGIETELDVV EGMQFDRGYL SQYMVTDNDK MEAVLENPYI LITDKKISNI QDILPLLEQI LQ QSRPLLI IADDVDGEAL PTLVLNKIRG TFNVVAVKAP GFGDRRKAML EDIAILTGGT VITDDLGLEL KDVTIENLGN ASK VVVDKD NTTIVEGSGE KEAIEARVQL IKNQIAETTS DFDREKLQER LAKLAGGVAV VKVGAATETE LKELKLRIED ALNA TRAAV EEGMVSGGGT ALVNVISKVS AVEAEGDVAT GIKIVVRALE EPIRQIAENA GYEGSVIVDK LKNVELGTGF NAATG EWVN MVEAGIVDPT KVTRSALQNA ASVSALLLTT EAVVADKPEP AAPAAPAMDP SMMGGMM UniProtKB: Chaperonin GroEL |
-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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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 62.5 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |