+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-40785 | |||||||||
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Title | MBP tagged mixed chain FAS, fusFAS_deltaACP | |||||||||
Map data | MBP tagged mixed chain FAS, fusFAS_deltaACP_corrected | |||||||||
Sample |
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Keywords | Fatty acid synthase / TRANSFERASE | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.87 Å | |||||||||
Authors | Mazhab-Jafari MT / Khalili Samani E | |||||||||
Funding support | Canada, 2 items
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Citation | Journal: Commun Biol / Year: 2024 Title: Direct structural analysis of a single acyl carrier protein domain in fatty acid synthase from the fungus Saccharomyces cerevisiae. Authors: Elnaz Khalili Samani / Amy C Chen / Jennifer W Lou / David L Dai / Alexander F A Keszei / Guihong Tan / Charles Boone / Martin Grininger / Mohammad T Mazhab-Jafari / Abstract: Acyl carrier protein (ACP) is the work horse of polyketide (PKS) and fatty acid synthases (FAS) and acts as a substrate shuttling domain in these mega enzymes. In fungi, FAS forms a 2.6 MDa symmetric ...Acyl carrier protein (ACP) is the work horse of polyketide (PKS) and fatty acid synthases (FAS) and acts as a substrate shuttling domain in these mega enzymes. In fungi, FAS forms a 2.6 MDa symmetric assembly with six identical copies of FAS1 and FAS2 polypeptides. However, ACP spatial distribution is not restricted by symmetry owing to the long and flexible loops that tether the shuttling domain to its corresponding FAS2 polypeptide. This symmetry breaking has hampered experimental investigation of substrate shuttling route in fungal FAS. Here, we develop a protein engineering and expression method to isolate asymmetric fungal FAS proteins containing odd numbers of ACP domains. Electron cryomicroscopy (cryoEM) observation of the engineered complex reveals a non-uniform distribution of the substrate shuttling domain relative to its corresponding FAS2 polypeptide at 2.9 Å resolution. This work lays the methodological foundation for experimental study of ACP shuttling route in fungi. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_40785.map.gz | 122.7 MB | EMDB map data format | |
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Header (meta data) | emd-40785-v30.xml emd-40785.xml | 14.3 KB 14.3 KB | Display Display | EMDB header |
Images | emd_40785.png | 90.3 KB | ||
Filedesc metadata | emd-40785.cif.gz | 4.2 KB | ||
Others | emd_40785_half_map_1.map.gz emd_40785_half_map_2.map.gz | 227 MB 227 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-40785 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-40785 | HTTPS FTP |
-Validation report
Summary document | emd_40785_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_40785_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_40785_validation.xml.gz | 16.2 KB | Display | |
Data in CIF | emd_40785_validation.cif.gz | 19.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-40785 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-40785 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_40785.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | MBP tagged mixed chain FAS, fusFAS_deltaACP_corrected | ||||||||||||||||||||||||||||||||||||
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:
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-Supplemental data
-Half map: Half map A
File | emd_40785_half_map_1.map | ||||||||||||
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Annotation | Half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map B
File | emd_40785_half_map_2.map | ||||||||||||
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Annotation | Half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : MBP-tagged mixed chain Fatty acid synthase, fusFAS_deltaACP
Entire | Name: MBP-tagged mixed chain Fatty acid synthase, fusFAS_deltaACP |
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Components |
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-Supramolecule #1: MBP-tagged mixed chain Fatty acid synthase, fusFAS_deltaACP
Supramolecule | Name: MBP-tagged mixed chain Fatty acid synthase, fusFAS_deltaACP type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Molecular weight | Theoretical: 2.7 MDa |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 2 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 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 25 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number grids imaged: 1 / Number real images: 15032 / Average electron dose: 36.0 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: 2.5 µm / Nominal defocus min: 0.6 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.87 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.0.2) / Number images used: 167630 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4) |
Final angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4) |