+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10748 | |||||||||
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Title | AP2 on a membrane containing tyrosine-based cargo peptide | |||||||||
Map data | Sharpened map of AP2 on membranes containing tyrosine-based cargo peptides | |||||||||
Sample |
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Keywords | clathrin / clathrin adaptor / ap2 / clathrin assembly / ENDOCYTOSIS | |||||||||
Function / homology | Function and homology information protein metabolic process => GO:0019538 / Gap junction degradation / Formation of annular gap junctions / LDL clearance / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / LDL clearance / WNT5A-dependent internalization of FZD4 / VLDLR internalisation and degradation / Retrograde neurotrophin signalling ...protein metabolic process => GO:0019538 / Gap junction degradation / Formation of annular gap junctions / LDL clearance / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / LDL clearance / WNT5A-dependent internalization of FZD4 / VLDLR internalisation and degradation / Retrograde neurotrophin signalling / Nef Mediated CD8 Down-regulation / Trafficking of GluR2-containing AMPA receptors / Retrograde neurotrophin signalling / Retrograde transport at the Trans-Golgi-Network / clathrin adaptor complex / WNT5A-dependent internalization of FZD4 / WNT5A-dependent internalization of FZD2, FZD5 and ROR2 / Trafficking of GluR2-containing AMPA receptors / extrinsic component of presynaptic endocytic zone membrane / WNT5A-dependent internalization of FZD4 / VLDLR internalisation and degradation / MHC class II antigen presentation / Recycling pathway of L1 / AP-2 adaptor complex / regulation of vesicle size / postsynaptic neurotransmitter receptor internalization / Recycling pathway of L1 / Cargo recognition for clathrin-mediated endocytosis / Retrograde neurotrophin signalling / Cargo recognition for clathrin-mediated endocytosis / positive regulation of synaptic vesicle endocytosis / clathrin-coated endocytic vesicle / LDL clearance / Clathrin-mediated endocytosis / clathrin adaptor activity / Clathrin-mediated endocytosis / membrane coat / vesicle budding from membrane / trans-Golgi network transport vesicle / clathrin-dependent endocytosis / MHC class II antigen presentation / membrane organization / coronary vasculature development / signal sequence binding / Nef Mediated CD4 Down-regulation / endolysosome membrane / aorta development / negative regulation of protein localization to plasma membrane / ventricular septum development / Neutrophil degranulation / low-density lipoprotein particle receptor binding / clathrin binding / Golgi Associated Vesicle Biogenesis / Recycling pathway of L1 / Trafficking of GluR2-containing AMPA receptors / positive regulation of receptor internalization / synaptic vesicle endocytosis / EPH-ephrin mediated repulsion of cells / post-translational protein modification / protein serine/threonine kinase binding / clathrin-coated pit / transport vesicle / vesicle-mediated transport / MHC class II antigen presentation / VLDLR internalisation and degradation / phosphatidylinositol binding / clathrin-coated endocytic vesicle membrane / kidney development / kinase binding / Post-translational protein phosphorylation / intracellular protein transport / trans-Golgi network / terminal bouton / receptor internalization / cytoplasmic side of plasma membrane / Regulation of Insulin-like Growth Factor (IGF) transport and uptake by Insulin-like Growth Factor Binding Proteins (IGFBPs) / Cargo recognition for clathrin-mediated endocytosis / disordered domain specific binding / endocytic vesicle membrane / Clathrin-mediated endocytosis / synaptic vesicle / presynapse / cytoplasmic vesicle / Potential therapeutics for SARS / protein-containing complex assembly / transmembrane transporter binding / postsynapse / membrane => GO:0016020 / endosome / protein domain specific binding / endoplasmic reticulum lumen / intracellular membrane-bounded organelle / lipid binding / glutamatergic synapse / synapse / protein-containing complex binding / protein kinase binding / Golgi apparatus / nucleoplasm / membrane Similarity search - Function | |||||||||
Biological species | Rattus norvegicus (Norway rat) / Homo sapiens (human) / Mus musculus (house mouse) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 9.1 Å | |||||||||
Authors | Kovtun O / Kane Dickson V | |||||||||
Funding support | United Kingdom, 2 items
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Citation | Journal: Sci Adv / Year: 2020 Title: Architecture of the AP2/clathrin coat on the membranes of clathrin-coated vesicles. Authors: Oleksiy Kovtun / Veronica Kane Dickson / Bernard T Kelly / David J Owen / John A G Briggs / Abstract: Clathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which ...Clathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which cargo-selecting clathrin adaptors are attached. Adaptor protein complex (AP2) is the key adaptor in CME. Crystallography has shown AP2 to adopt a range of conformations. Here, we used cryo-electron microscopy, tomography, and subtomogram averaging to determine structures, interactions, and arrangements of clathrin and AP2 at the key steps of coat assembly, from AP2 in solution to membrane-assembled clathrin-coated vesicles (CCVs). AP2 binds cargo and PtdIns(4,5) (phosphatidylinositol 4,5-bisphosphate)-containing membranes via multiple interfaces, undergoing conformational rearrangement from its cytosolic state. The binding mode of AP2 β2 appendage into the clathrin lattice in CCVs and buds implies how the adaptor structurally modulates coat curvature and coat disassembly. | |||||||||
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_10748.map.gz | 1.6 MB | EMDB map data format | |
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Header (meta data) | emd-10748-v30.xml emd-10748.xml | 28.5 KB 28.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_10748_fsc.xml | 4.7 KB | Display | FSC data file |
Images | emd_10748.png | 231.9 KB | ||
Masks | emd_10748_msk_1.map | 8 MB | Mask map | |
Filedesc metadata | emd-10748.cif.gz | 8.2 KB | ||
Others | emd_10748_half_map_1.map.gz emd_10748_half_map_2.map.gz | 7.4 MB 7.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10748 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10748 | HTTPS FTP |
-Validation report
Summary document | emd_10748_validation.pdf.gz | 674.8 KB | Display | EMDB validaton report |
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Full document | emd_10748_full_validation.pdf.gz | 674.4 KB | Display | |
Data in XML | emd_10748_validation.xml.gz | 10.4 KB | Display | |
Data in CIF | emd_10748_validation.cif.gz | 13.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10748 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10748 | HTTPS FTP |
-Related structure data
Related structure data | 6yafMC 6yaeC 6yahC 6yaiC C: citing same article (ref.) M: atomic model generated by this map |
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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_10748.map.gz / Format: CCP4 / Size: 8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Sharpened map of AP2 on membranes containing tyrosine-based cargo peptides | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.78 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_10748_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: Unsharpened half-map of AP2 on membranes containing tyrosine-based...
File | emd_10748_half_map_1.map | ||||||||||||
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Annotation | Unsharpened half-map of AP2 on membranes containing tyrosine-based cargo peptides | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unsharpened half-map of AP2 on membranes containing tyrosine-based...
File | emd_10748_half_map_2.map | ||||||||||||
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Annotation | Unsharpened half-map of AP2 on membranes containing tyrosine-based cargo peptides | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : AP2 on membranes
+Supramolecule #1: AP2 on membranes
+Supramolecule #2: AP-2 complex subunits alpha-2 and mu
+Supramolecule #3: AP-2 complex subunit beta
+Supramolecule #4: AP-2 complex subunit sigma
+Supramolecule #5: Tyrosine type cargo signal derived from TGN38
+Macromolecule #1: AP-2 complex subunit alpha-2
+Macromolecule #2: AP-2 complex subunit beta
+Macromolecule #3: AP-2 complex subunit mu
+Macromolecule #4: Tyrosine type cargo signal derived from TGN38
+Macromolecule #5: AP-2 complex subunit sigma
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | 3D array |
-Sample preparation
Concentration | 0.3 mg/mL | ||||||||||
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Buffer | pH: 7.2 Component:
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Grid | Model: C-flat-2/2 / Support film - Material: CARBON / Support film - topology: HOLEY | ||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 98 % / Chamber temperature: 291 K / Instrument: LEICA EM GP Details: The sample was supplemented with 10 nm nanogold fiducials, and 3 ul of the mixture was backside blotted for 3 seconds.. | ||||||||||
Details | The sample contained AP2 and 400 nm extruded liposomes |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-10 / Number grids imaged: 2 / Average exposure time: 0.2 sec. / Average electron dose: 3.2 e/Å2 Details: The images were collected in movie mode at 10 frames per second |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 6.5 µm / Calibrated defocus min: 1.5 µm / Calibrated magnification: 81000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 6.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |