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- EMDB-2897: Cryo-EM structure of helical ANTH and ENTH tubules on PI(4,5)P2-c... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2897 | |||||||||
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Title | Cryo-EM structure of helical ANTH and ENTH tubules on PI(4,5)P2-containing membranes | |||||||||
![]() | Reconstruction of the one-start helix, of pitch 63.2 Angstrom with 9.96 subunits per turn, formed by ANTH-ENTH assembly on PIP2 containing GUVs. | |||||||||
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![]() | epsin / Hip1R / ENTH / ANTH / clathrin adaptors / endocytosis / PI(4 / 5)P2 | |||||||||
Function / homology | ![]() Cargo recognition for clathrin-mediated endocytosis / actin cortical patch assembly / clathrin vesicle coat / clathrin light chain binding / negative regulation of Arp2/3 complex-mediated actin nucleation / actin cortical patch / incipient cellular bud site / cellular bud tip / clathrin coat assembly / clathrin adaptor activity ...Cargo recognition for clathrin-mediated endocytosis / actin cortical patch assembly / clathrin vesicle coat / clathrin light chain binding / negative regulation of Arp2/3 complex-mediated actin nucleation / actin cortical patch / incipient cellular bud site / cellular bud tip / clathrin coat assembly / clathrin adaptor activity / cellular bud neck / mating projection tip / phosphatidylinositol-3,4-bisphosphate binding / phosphatidylinositol-3,5-bisphosphate binding / clathrin-coated vesicle / clathrin binding / ubiquitin binding / actin filament organization / phospholipid binding / endocytosis / actin filament binding / early endosome / endosome / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 18.3 Å | |||||||||
![]() | Skruzny M / Desfosses A / Prinz S / Dodonova SO / Gieras A / Uetrecht C / Jakobi AJ / Abella M / Hagen WJH / Schulz J ...Skruzny M / Desfosses A / Prinz S / Dodonova SO / Gieras A / Uetrecht C / Jakobi AJ / Abella M / Hagen WJH / Schulz J / Meijers R / Rybin V / Briggs JAG / Sachse C / Kaksonen M | |||||||||
![]() | ![]() Title: An organized co-assembly of clathrin adaptors is essential for endocytosis. Authors: Michal Skruzny / Ambroise Desfosses / Simone Prinz / Svetlana O Dodonova / Anna Gieras / Charlotte Uetrecht / Arjen J Jakobi / Marc Abella / Wim J H Hagen / Joachim Schulz / Rob Meijers / ...Authors: Michal Skruzny / Ambroise Desfosses / Simone Prinz / Svetlana O Dodonova / Anna Gieras / Charlotte Uetrecht / Arjen J Jakobi / Marc Abella / Wim J H Hagen / Joachim Schulz / Rob Meijers / Vladimir Rybin / John A G Briggs / Carsten Sachse / Marko Kaksonen / ![]() Abstract: Clathrin-mediated endocytosis, the main trafficking route from the plasma membrane to the cytoplasm, is critical to many fundamental cellular processes. Clathrin, coupled to the membrane by adaptor ...Clathrin-mediated endocytosis, the main trafficking route from the plasma membrane to the cytoplasm, is critical to many fundamental cellular processes. Clathrin, coupled to the membrane by adaptor proteins, is thought to play a major structural role in endocytosis by self-assembling into a cage-like lattice around the forming vesicle. Although clathrin adaptors are essential for endocytosis, little is known about their structural role in this process. Here we show that the membrane-binding domains of two conserved clathrin adaptors, Sla2 and Ent1, co-assemble in a PI(4,5)P2-dependent manner to form organized lattices on membranes. We determined the structure of the co-assembled lattice by electron cryo-microscopy and designed mutations that specifically impair the lattice formation in vitro. We show that these mutations block endocytosis in vivo. We suggest that clathrin adaptors not only link the polymerized clathrin to the membrane but also form an oligomeric structure, which is essential for membrane remodeling during endocytosis. | |||||||||
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 | ![]() | 52.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 11.7 KB 11.7 KB | Display Display | ![]() |
Images | ![]() | 158.5 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 224.8 KB | Display | ![]() |
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Full document | ![]() | 224 KB | Display | |
Data in XML | ![]() | 4.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Reconstruction of the one-start helix, of pitch 63.2 Angstrom with 9.96 subunits per turn, formed by ANTH-ENTH assembly on PIP2 containing GUVs. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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
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Sample components
-Entire : ANTH and ENTH domains of the clathrin adaptors Sla2 and Ent1, res...
Entire | Name: ANTH and ENTH domains of the clathrin adaptors Sla2 and Ent1, respectively. |
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Components |
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-Supramolecule #1000: ANTH and ENTH domains of the clathrin adaptors Sla2 and Ent1, res...
Supramolecule | Name: ANTH and ENTH domains of the clathrin adaptors Sla2 and Ent1, respectively. type: sample / ID: 1000 / Number unique components: 2 |
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-Macromolecule #1: ANTH domain of endocytic adaptor Sla2
Macromolecule | Name: ANTH domain of endocytic adaptor Sla2 / type: protein_or_peptide / ID: 1 / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Experimental: 33.215 KDa / Theoretical: 33.21 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Protein SLA2 / InterPro: AP180 N-terminal homology (ANTH) domain |
-Macromolecule #2: ENTH domain of epsin Ent1
Macromolecule | Name: ENTH domain of epsin Ent1 / type: protein_or_peptide / ID: 2 / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Experimental: 18.87 KDa / Theoretical: 18.847 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Epsin-1 / InterPro: ENTH domain |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | helical array |
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Sample preparation
Buffer | pH: 7.5 / Details: 20 mM HEPES, pH 7.5; 100 mM KCl |
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Grid | Details: on C-flat holey carbon coated grids (Protochips) |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 80 % / Instrument: HOMEMADE PLUNGER Method: applied on C-flat holey carbon coated grids (Protochips) and vitrified |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Date | Mar 1, 2013 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 1419 / Average electron dose: 10 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 59000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 5.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 59000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Details | Processing done with SPRING |
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CTF correction | Details: Each Particle |
Final reconstruction | Applied symmetry - Helical parameters - Δz: 6.345 Å Applied symmetry - Helical parameters - Δ&Phi: 36.145 ° Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric) Resolution.type: BY AUTHOR / Resolution: 18.3 Å / Resolution method: OTHER / Software - Name: SPRING / Number images used: 154432 |