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- EMDB-2411: Hsc70-induced Changes in Clathrin-Auxilin Cage Structure Suggest ... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2411 | |||||||||
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Title | Hsc70-induced Changes in Clathrin-Auxilin Cage Structure Suggest a Role for Clathrin Light Chains in Cage Disassembly | |||||||||
![]() | Map of Clathrin-Auxilin complex. Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms. | |||||||||
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![]() | Endocytosis / coated vesicles / clathrin / auxilin | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 31.0 Å | |||||||||
![]() | Young A / Stoilova-McPhie S / Rothnie A / Vallis Y / Harvey-Smith P / Ranson N / Kent H / Brodsky FM / Pearse BM / Roseman A / Smith CJ | |||||||||
![]() | Journal: J Mol Biol / Year: 2004 Title: Location of auxilin within a clathrin cage. Authors: Corinne J Smith / Timothy R Dafforn / Helen Kent / Catherine A Sims / Kavita Khubchandani-Aswani / Lin Zhang / Helen R Saibil / Barbara M F Pearse / ![]() Abstract: The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism ...The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism but until now there was no structural information on how auxilin interacts with the clathrin cage. Here we have determined the three-dimensional structure of a complex of auxilin with clathrin cages by cryo-electron microscopy and single particle analysis. We show that auxilin forms a discrete shell of density on the inside of the clathrin cage. Peptide competition assays confirm that a candidate clathrin box motif in auxilin, LLGLE, can bind to a clathrin construct containing the beta-propeller domain and also displace the well-characterised LLNLD clathrin box motif derived from the beta-adaptin hinge region. The means by which auxilin could both aid clathrin coat assembly and displace clathrin from AP2 during uncoating is discussed. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 58.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10.2 KB 10.2 KB | Display Display | ![]() |
Images | ![]() | 530.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Map of Clathrin-Auxilin complex. Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 6.4 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Clathrin bound to auxilin
Entire | Name: Clathrin bound to auxilin |
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Components |
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-Supramolecule #1000: Clathrin bound to auxilin
Supramolecule | Name: Clathrin bound to auxilin / type: sample / ID: 1000 / Number unique components: 2 |
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-Macromolecule #1: Clathrin
Macromolecule | Name: Clathrin / type: protein_or_peptide / ID: 1 / Recombinant expression: No |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #2: Auxilin
Macromolecule | Name: Auxilin / type: protein_or_peptide / ID: 2 / Recombinant expression: No |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 6 Details: 20 mM Mes (pH 6.0), 2 mM magnesium acetate, 25 mM KCl, 10 mM (NH4)2SO4, 1 mM DTT |
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Vitrification | Cryogen name: ETHANE / Instrument: HOMEMADE PLUNGER |
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Electron microscopy #1
Microscopy ID | 1 |
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Microscope | FEI/PHILIPS CM200FEG |
Date | Feb 2, 2000 |
Image recording | Digitization - Scanner: ZEISS SCAI |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: GATAN LIQUID NITROGEN |
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Electron microscopy #2
Microscopy ID | 2 |
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Microscope | FEI/PHILIPS CM200FEG |
Date | Feb 23, 2000 |
Image recording | Digitization - Scanner: ZEISS SCAI |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: GATAN LIQUID NITROGEN |
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Image processing
CTF correction | Details: Whole micrographs |
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Final reconstruction | Applied symmetry - Point group: D6 (2x6 fold dihedral) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 31.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: MRC, SPIDER, FREALIGN Details: Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms. Number images used: 1745 |