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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-21325 | |||||||||
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| Title | Focused Classification of FIP200 NTD | |||||||||
Map data | Final FIP200 NTD Map | |||||||||
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| Function / homology | Function and homology informationAtg1/ULK1 kinase complex / ribophagy / glycophagy / phagophore assembly site membrane / pexophagy / autophagy of mitochondrion / piecemeal microautophagy of the nucleus / phagophore assembly site / reticulophagy / Macroautophagy ...Atg1/ULK1 kinase complex / ribophagy / glycophagy / phagophore assembly site membrane / pexophagy / autophagy of mitochondrion / piecemeal microautophagy of the nucleus / phagophore assembly site / reticulophagy / Macroautophagy / autophagosome membrane / autophagosome assembly / positive regulation of cell size / extrinsic apoptotic signaling pathway / protein-membrane adaptor activity / positive regulation of autophagy / negative regulation of extrinsic apoptotic signaling pathway / positive regulation of JNK cascade / liver development / autophagy / heart development / nuclear membrane / defense response to virus / molecular adaptor activity / lysosome / negative regulation of cell population proliferation / innate immune response / endoplasmic reticulum membrane / protein kinase binding / cytosol Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 8.4 Å | |||||||||
Authors | Yokom AL / Shi X / Hurley JH | |||||||||
Citation | Journal: J Cell Biol / Year: 2020Title: ULK complex organization in autophagy by a C-shaped FIP200 N-terminal domain dimer. Authors: Xiaoshan Shi / Adam L Yokom / Chunxin Wang / Lindsey N Young / Richard J Youle / James H Hurley / ![]() Abstract: The autophagy-initiating human ULK complex consists of the kinase ULK1/2, FIP200, ATG13, and ATG101. Hydrogen-deuterium exchange mass spectrometry was used to map their mutual interactions. The N- ...The autophagy-initiating human ULK complex consists of the kinase ULK1/2, FIP200, ATG13, and ATG101. Hydrogen-deuterium exchange mass spectrometry was used to map their mutual interactions. The N-terminal 640 residues (NTD) of FIP200 interact with the C-terminal IDR of ATG13. Mutations in these regions abolish their interaction. Negative stain EM and multiangle light scattering showed that FIP200 is a dimer, while a single molecule each of the other subunits is present. The FIP200NTD is flexible in the absence of ATG13, but in its presence adopts the shape of the letter C ∼20 nm across. The ULK1 EAT domain interacts loosely with the NTD dimer, while the ATG13:ATG101 HORMA dimer does not contact the NTD. Cryo-EM of the NTD dimer revealed a structural similarity to the scaffold domain of TBK1, suggesting an evolutionary similarity between the autophagy-initiating TBK1 kinase and the ULK1 kinase complex. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_21325.map.gz | 111.6 KB | EMDB map data format | |
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| Header (meta data) | emd-21325-v30.xml emd-21325.xml | 12.1 KB 12.1 KB | Display Display | EMDB header |
| Images | emd_21325.png | 34.5 KB | ||
| Others | emd_21325_half_map_1.map.gz emd_21325_half_map_2.map.gz | 20.7 MB 20.7 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21325 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21325 | HTTPS FTP |
-Validation report
| Summary document | emd_21325_validation.pdf.gz | 78.9 KB | Display | EMDB validaton report |
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| Full document | emd_21325_full_validation.pdf.gz | 78 KB | Display | |
| Data in XML | emd_21325_validation.xml.gz | 494 B | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21325 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21325 | HTTPS FTP |
-Related structure data
| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_21325.map.gz / Format: CCP4 / Size: 22.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | Final FIP200 NTD Map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 2.274 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: Half Map B
| File | emd_21325_half_map_1.map | ||||||||||||
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| Annotation | Half Map B | ||||||||||||
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| Density Histograms |
-Half map: Half Map A
| File | emd_21325_half_map_2.map | ||||||||||||
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| Annotation | Half Map A | ||||||||||||
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| Density Histograms |
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Sample components
-Entire : FIP200 NTD Focused Classification Monomer
| Entire | Name: FIP200 NTD Focused Classification Monomer |
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| Components |
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-Supramolecule #1: FIP200 NTD Focused Classification Monomer
| Supramolecule | Name: FIP200 NTD Focused Classification Monomer / type: complex / ID: 1 / Parent: 0 |
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| Source (natural) | Organism: Homo sapiens (human) |
| Recombinant expression | Organism: Homo sapiens (human) / Recombinant strain: HEK293T |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.5 |
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| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 20 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | FEI TALOS ARCTICA |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 60.0 e/Å2 |
| Electron beam | Acceleration voltage: 200 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 |
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
| CTF correction | Software - Name: CTFFIND (ver. 4) |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 8.4 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 29198 |
| Initial angle assignment | Type: RANDOM ASSIGNMENT |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
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