[English] 日本語
![](img/lk-miru.gif)
- EMDB-2532: Tomographic subvolume average of EFF-1 fusogen on extracellular v... -
+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-2532 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Tomographic subvolume average of EFF-1 fusogen on extracellular vesicles | |||||||||
![]() | Tomographic subvolume average of EFF-1 on nanotubular extracellular vesicles | |||||||||
![]() |
| |||||||||
![]() | cell-cell fusion / extracellular fusion / membrane fusion / fusogen / pre-fusion state | |||||||||
Function / homology | ![]() nematode male tail mating organ morphogenesis / fusogenic activity / nematode pharyngeal muscle development / EFF-1 complex / post-embryonic body morphogenesis / nematode male tail tip morphogenesis / plasma membrane fusion / vulval development / cell-cell fusion / syncytium formation by plasma membrane fusion ...nematode male tail mating organ morphogenesis / fusogenic activity / nematode pharyngeal muscle development / EFF-1 complex / post-embryonic body morphogenesis / nematode male tail tip morphogenesis / plasma membrane fusion / vulval development / cell-cell fusion / syncytium formation by plasma membrane fusion / embryonic body morphogenesis / egg-laying behavior / cell-cell contact zone / locomotion / plasma membrane => GO:0005886 / morphogenesis of an epithelium / kinase activity / phosphorylation / extracellular region / identical protein binding / plasma membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 45.0 Å | |||||||||
![]() | Zeev-Ben-Mordehai T / Vasishtan D / Siebert CA / Grunewald K | |||||||||
![]() | ![]() Title: The full-length cell-cell fusogen EFF-1 is monomeric and upright on the membrane. Authors: Tzviya Zeev-Ben-Mordehai / Daven Vasishtan / C Alistair Siebert / Kay Grünewald / ![]() Abstract: Fusogens are membrane proteins that remodel lipid bilayers to facilitate membrane merging. Although several fusogen ectodomain structures have been solved, structural information on full-length, ...Fusogens are membrane proteins that remodel lipid bilayers to facilitate membrane merging. Although several fusogen ectodomain structures have been solved, structural information on full-length, natively membrane-anchored fusogens is scarce. Here we present the electron cryo microscopy three-dimensional reconstruction of the Caenorhabditis elegans epithelial fusion failure 1 (EFF-1) protein natively anchored in cell-derived membrane vesicles. This reveals a membrane protruding, asymmetric, elongated monomer. Flexible fitting of a protomer of the EFF-1 crystal structure, which is homologous to viral class-II fusion proteins, shows that EFF-1 has a hairpin monomeric conformation before fusion. These structural insights, when combined with our observations of membrane-merging intermediates between vesicles, enable us to propose a model for EFF-1 mediated fusion. This process, involving identical proteins on both membranes to be fused, follows a mechanism that shares features of SNARE-mediated fusion while using the structural building blocks of the unilaterally acting class-II viral fusion proteins. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 2.5 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 10.7 KB 10.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 3.9 KB | Display | ![]() |
Images | ![]() | 65.6 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 234.2 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 233.4 KB | Display | |
Data in XML | ![]() | 7.3 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Tomographic subvolume average of EFF-1 on nanotubular extracellular vesicles | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 3.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : Epithelial fusion failure 1 (EFF-1) Isoform A on extracellular ve...
Entire | Name: Epithelial fusion failure 1 (EFF-1) Isoform A on extracellular vesicles |
---|---|
Components |
|
-Supramolecule #1000: Epithelial fusion failure 1 (EFF-1) Isoform A on extracellular ve...
Supramolecule | Name: Epithelial fusion failure 1 (EFF-1) Isoform A on extracellular vesicles type: sample / ID: 1000 / Oligomeric state: Monomer / Number unique components: 1 |
---|---|
Molecular weight | Theoretical: 87 KDa |
-Macromolecule #1: Epithelial fusion failure 1, isoform a
Macromolecule | Name: Epithelial fusion failure 1, isoform a / type: protein_or_peptide / ID: 1 / Name.synonym: EFF-1a / Details: Proteins on extracellular vesicles / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 87 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: EFF-1A GO: plasma membrane fusion, syncytium formation by plasma membrane fusion, plasma membrane => GO:0005886, extracellular region, identical protein binding |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | subtomogram averaging |
Aggregation state | particle |
-
Sample preparation
Buffer | pH: 7.4 / Details: 25mM HEPES, 130mM NaCl |
---|---|
Grid | Details: Holey carbon on top of 200 mesh gold grid. |
Vitrification | Cryogen name: ETHANE-PROPANE MIXTURE / Chamber temperature: 77 K / Instrument: HOMEMADE PLUNGER / Method: Blot for 3 seconds before plunging |
-
Electron microscopy
Microscope | FEI POLARA 300 |
---|---|
Temperature | Min: 80 K / Max: 100 K / Average: 85 K |
Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 115,000 times magnification Legacy - Electron beam tilt params: +1 to -1 mradians |
Specialist optics | Energy filter - Name: Gatan Quantum 964 / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV |
Date | Mar 3, 2013 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Digitization - Sampling interval: 30 µm / Number real images: 39 / Average electron dose: 60 e/Å2 / Details: 1 tomogram created from 39 projection images / Bits/pixel: 16 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 78950 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 2.0 µm / Nominal magnification: 95000 |
Sample stage | Specimen holder: Liquid nitrogen cooled / Specimen holder model: GATAN HELIUM / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° |
Experimental equipment | ![]() Model: Tecnai Polara / Image courtesy: FEI Company |