+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-23763 | |||||||||||||||||||||||||||
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Title | The F1 region of ammocidin-bound Saccharomyces cerevisiae ATP synthase | |||||||||||||||||||||||||||
Map data | Locally sharpened map. | |||||||||||||||||||||||||||
Sample |
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Keywords | F1-ATPase / inhibitor / anti-cancer / HYDROLASE / HYDROLASE-HYDROLASE INHIBITOR complex | |||||||||||||||||||||||||||
Function / homology | Function and homology information : / : / : / : / proton motive force-driven ATP synthesis / proton motive force-driven mitochondrial ATP synthesis / proton-transporting ATP synthase complex, catalytic core F(1) / H+-transporting two-sector ATPase / proton-transporting ATPase activity, rotational mechanism / proton-transporting ATP synthase activity, rotational mechanism ...: / : / : / : / proton motive force-driven ATP synthesis / proton motive force-driven mitochondrial ATP synthesis / proton-transporting ATP synthase complex, catalytic core F(1) / H+-transporting two-sector ATPase / proton-transporting ATPase activity, rotational mechanism / proton-transporting ATP synthase activity, rotational mechanism / ADP binding / mitochondrial inner membrane / ATP hydrolysis activity / mitochondrion / ATP binding Similarity search - Function | |||||||||||||||||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||||||||||||||||||||
Authors | Guo H / Rubinstein JL | |||||||||||||||||||||||||||
Funding support | Canada, United States, 8 items
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Citation | Journal: Nat Chem Biol / Year: 2022 Title: Apoptolidin family glycomacrolides target leukemia through inhibition of ATP synthase. Authors: Benjamin J Reisman / Hui Guo / Haley E Ramsey / Madison T Wright / Bradley I Reinfeld / P Brent Ferrell / Gary A Sulikowski / W Kimryn Rathmell / Michael R Savona / Lars Plate / John L ...Authors: Benjamin J Reisman / Hui Guo / Haley E Ramsey / Madison T Wright / Bradley I Reinfeld / P Brent Ferrell / Gary A Sulikowski / W Kimryn Rathmell / Michael R Savona / Lars Plate / John L Rubinstein / Brian O Bachmann / Abstract: Cancer cells have long been recognized to exhibit unique bioenergetic requirements. The apoptolidin family of glycomacrolides are distinguished by their selective cytotoxicity towards oncogene- ...Cancer cells have long been recognized to exhibit unique bioenergetic requirements. The apoptolidin family of glycomacrolides are distinguished by their selective cytotoxicity towards oncogene-transformed cells, yet their molecular mechanism remains uncertain. We used photoaffinity analogs of the apoptolidins to identify the F subcomplex of mitochondrial ATP synthase as the target of apoptolidin A. Cryogenic electron microscopy (cryo-EM) of apoptolidin and ammocidin-ATP synthase complexes revealed a novel shared mode of inhibition that was confirmed by deep mutational scanning of the binding interface to reveal resistance mutations which were confirmed using CRISPR-Cas9. Ammocidin A was found to suppress leukemia progression in vivo at doses that were tolerated with minimal toxicity. The combination of cellular, structural, mutagenesis, and in vivo evidence defines the mechanism of action of apoptolidin family glycomacrolides and establishes a path to address oxidative phosphorylation-dependent cancers. | |||||||||||||||||||||||||||
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_23763.map.gz | 110.6 MB | EMDB map data format | |
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Header (meta data) | emd-23763-v30.xml emd-23763.xml | 27.1 KB 27.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_23763_fsc.xml | 11.4 KB | Display | FSC data file |
Images | emd_23763.png | 87.5 KB | ||
Masks | emd_23763_msk_1.map | 125 MB | Mask map | |
Filedesc metadata | emd-23763.cif.gz | 7.2 KB | ||
Others | emd_23763_additional_1.map.gz emd_23763_half_map_1.map.gz emd_23763_half_map_2.map.gz | 62.6 MB 116 MB 116 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-23763 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23763 | HTTPS FTP |
-Validation report
Summary document | emd_23763_validation.pdf.gz | 863.4 KB | Display | EMDB validaton report |
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Full document | emd_23763_full_validation.pdf.gz | 863 KB | Display | |
Data in XML | emd_23763_validation.xml.gz | 18.5 KB | Display | |
Data in CIF | emd_23763_validation.cif.gz | 24.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23763 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23763 | HTTPS FTP |
-Related structure data
Related structure data | 7md2MC 7md3C M: atomic model generated by this map C: citing same article (ref.) |
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Similar 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_23763.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Locally sharpened map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.03 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_23763_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Unsharpened map.
File | emd_23763_additional_1.map | ||||||||||||
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Annotation | Unsharpened map. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1.
File | emd_23763_half_map_1.map | ||||||||||||
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Annotation | Half map 1. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2.
File | emd_23763_half_map_2.map | ||||||||||||
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Annotation | Half map 2. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : The F1 region of ammocidin-bound Saccharomyces cerevisiae ATP synthase
Entire | Name: The F1 region of ammocidin-bound Saccharomyces cerevisiae ATP synthase |
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Components |
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-Supramolecule #1: The F1 region of ammocidin-bound Saccharomyces cerevisiae ATP synthase
Supramolecule | Name: The F1 region of ammocidin-bound Saccharomyces cerevisiae ATP synthase type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4 |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: USY006 |
Molecular weight | Theoretical: 400 KDa |
-Macromolecule #1: ATP synthase subunit alpha
Macromolecule | Name: ATP synthase subunit alpha / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: USY006 |
Molecular weight | Theoretical: 55.007402 KDa |
Sequence | String: ASTKAQPTEV SSILEERIKG VSDEANLNET GRVLAVGDGI ARVFGLNNIQ AEELVEFSSG VKGMALNLEP GQVGIVLFGS DRLVKEGEL VKRTGNIVDV PVGPGLLGRV VDALGNPIDG KGPIDAAGRS RAQVKAPGIL PRRSVHEPVQ TGLKAVDALV P IGRGQREL ...String: ASTKAQPTEV SSILEERIKG VSDEANLNET GRVLAVGDGI ARVFGLNNIQ AEELVEFSSG VKGMALNLEP GQVGIVLFGS DRLVKEGEL VKRTGNIVDV PVGPGLLGRV VDALGNPIDG KGPIDAAGRS RAQVKAPGIL PRRSVHEPVQ TGLKAVDALV P IGRGQREL IIGDRQTGKT AVALDTILNQ KRWNNGSDES KKLYCVYVAV GQKRSTVAQL VQTLEQHDAM KYSIIVAATA SE AAPLQYL APFTAASIGE WFRDNGKHAL IVYDDLSKQA VAYRQLSLLL RRPPGREAYP GDVFYLHSRL LERAAKLSEK EGS GSLTAL PVIETQGGDV SAYIPTNVIS ITDGQIFLEA ELFYKGIRPA INVGLSVSRV GSAAQVKALK QVAGSLKLFL AQYR EVAAF AQFGSDLDAS TKQTLVRGER LTQLLKQNQY SPLATEEQVP LIYAGVNGHL DGIELSRIGE FESSFLSYLK SNHNE LLTE IREKGELSKE LLASLKSATE SFVATF UniProtKB: ATP synthase subunit alpha |
-Macromolecule #2: ATP synthase subunit beta
Macromolecule | Name: ATP synthase subunit beta / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO / EC number: H+-transporting two-sector ATPase |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: USY006 |
Molecular weight | Theoretical: 51.181082 KDa |
Sequence | String: ASAAQSTPIT GKVTAVIGAI VDVHFEQSEL PAILNALEIK TPQGKLVLEV AQHLGENTVR TIAMDGTEGL VRGEKVLDTG GPISVPVGR ETLGRIINVI GEPIDERGPI KSKLRKPIHA DPPSFAEQST SAEILETGIK VVDLLAPYAR GGKIGLFGGA G VGKTVFIQ ...String: ASAAQSTPIT GKVTAVIGAI VDVHFEQSEL PAILNALEIK TPQGKLVLEV AQHLGENTVR TIAMDGTEGL VRGEKVLDTG GPISVPVGR ETLGRIINVI GEPIDERGPI KSKLRKPIHA DPPSFAEQST SAEILETGIK VVDLLAPYAR GGKIGLFGGA G VGKTVFIQ ELINNIAKAH GGFSVFTGVG ERTREGNDLY REMKETGVIN LEGESKVALV FGQMNEPPGA RARVALTGLT IA EYFRDEE GQDVLLFIDN IFRFTQAGSE VSALLGRIPS AVGYQPTLAT DMGLLQERIT TTKKGSVTSV QAVYVPADDL TDP APATTF AHLDATTVLS RGISELGIYP AVDPLDSKSR LLDAAVVGQE HYDVASKVQE TLQTYKSLQD IIAILGMDEL SEQD KLTVE RARKIQRFLS QPFAVAEVFT GIPGKLVRLK DTVASFKAVL EGKYDNIPEH AFYMVGGIED VVAKAEKLAA EAN UniProtKB: ATP synthase subunit beta |
-Macromolecule #3: ATP synthase subunit gamma
Macromolecule | Name: ATP synthase subunit gamma / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: USY006 |
Molecular weight | Theoretical: 30.65716 KDa |
Sequence | String: ATLKEVEMRL KSIKNIEKIT KTMKIVASTR LSKAEKAKIS AKKMDEAEQL FYKNAETKNL DVEATETGAP KELIVAITSD KGLCGSIHS QLAKAVRRHL NDQPNADIVT IGDKIKMQLL RTHPNNIKLS INGIGKDAPT FQESALIADK LLSVMKAGTY P KISIFYND ...String: ATLKEVEMRL KSIKNIEKIT KTMKIVASTR LSKAEKAKIS AKKMDEAEQL FYKNAETKNL DVEATETGAP KELIVAITSD KGLCGSIHS QLAKAVRRHL NDQPNADIVT IGDKIKMQLL RTHPNNIKLS INGIGKDAPT FQESALIADK LLSVMKAGTY P KISIFYND PVSSLSFEPS EKPIFNAKTI EQSPSFGKFE IDTDANVPRD LFEYTLANQM LTAMAQGYAA EISARRNAMD NA SKNAGDM INRYSILYNR TRQAVITNEL VDIITGASSL G UniProtKB: ATP synthase subunit gamma |
-Macromolecule #4: ATP synthase subunit epsilon, mitochondrial
Macromolecule | Name: ATP synthase subunit epsilon, mitochondrial / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: USY006 |
Molecular weight | Theoretical: 6.618359 KDa |
Sequence | String: SAWRKAGISY AAYLNVAAQA IRSSLKTELQ TASVLNRSQT DAFYTQYKNG TAASEPTPIT K UniProtKB: ATP synthase subunit epsilon, mitochondrial |
-Macromolecule #5: ADENOSINE-5'-TRIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 5 / Number of copies: 3 / Formula: ATP |
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Molecular weight | Theoretical: 507.181 Da |
Chemical component information | ChemComp-ATP: |
-Macromolecule #6: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 6 / Number of copies: 3 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Macromolecule #7: PHOSPHATE ION
Macromolecule | Name: PHOSPHATE ION / type: ligand / ID: 7 / Number of copies: 1 / Formula: PO4 |
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Molecular weight | Theoretical: 94.971 Da |
Chemical component information | ChemComp-PO4: |
-Macromolecule #8: (3~{E},5~{Z},7~{E},9~{R},10~{S},11~{E},13~{E},15~{E},17~{R},18~{S...
Macromolecule | Name: (3~{E},5~{Z},7~{E},9~{R},10~{S},11~{E},13~{E},15~{E},17~{R},18~{S},20~{S})-20-[(1~{R})-1-[(2~{S},3~{R},4~{R},5~{S},6~{R})-5-[(2~{S},4~{S},5~{S},6~{R})-5-[(2~{S},4~{R},5~{R},6~{R})-4,6-dimethyl- ...Name: (3~{E},5~{Z},7~{E},9~{R},10~{S},11~{E},13~{E},15~{E},17~{R},18~{S},20~{S})-20-[(1~{R})-1-[(2~{S},3~{R},4~{R},5~{S},6~{R})-5-[(2~{S},4~{S},5~{S},6~{R})-5-[(2~{S},4~{R},5~{R},6~{R})-4,6-dimethyl-4,5-bis(oxidanyl)oxan-2-yl]oxy-6-methyl-4-oxidanyl-oxan-2-yl]oxy-3-methoxy-6-(3-methoxypropyl)-5-methyl-2,4-bis(oxidanyl)oxan-2-yl]ethyl]-5,18-dimethoxy-3,7,9,11,13,15-hexamethyl-10-[(2~{R},3~{S},4~{R},5~{R},6~{S})-6-methyl-3,4,5-tris(oxidanyl)oxan-2-yl]oxy-17-oxidanyl-1-oxacycloicosa-3,5,7,11,13,15-hexaen-2-one type: ligand / ID: 8 / Number of copies: 1 / Formula: ZHD |
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Molecular weight | Theoretical: 1.15738 KDa |
Chemical component information | ChemComp-ZHD: |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 15 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: Homemade / Material: COPPER/RHODIUM / Mesh: 400 / Support film - Material: GOLD / Support film - topology: HOLEY / Support film - Film thickness: 30 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 120 sec. |
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 4 K / Instrument: FEI VITROBOT MARK III |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Number real images: 4345 / Average exposure time: 9.5 sec. / Average electron dose: 45.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated defocus max: 2.4 µm / Calibrated magnification: 135922 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus min: 0.7000000000000001 µm / Nominal magnification: 75000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: RECIPROCAL / Protocol: AB INITIO MODEL |
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Output model | PDB-7md2: |