EMD-2415 Experiments and Validation

Helical reconstruction of HMPV matrix protein-lipid filaments

Helical reconstruction
Overview of EMD-2415
Sample name: Human metapneumovirus matrix protein M bound to DOPC
Organisms: Human metapneumovirus, synthetic construct

Map parameters

Recommended contour level: 2.00 (author)
Number of grid points: 256 × 256 × 256
Voxel size: 3.10 × 3.10 × 3.10 Å
Minimum density: -6.341
Maximum density: 10.959
Average density: 0.026
Standard deviation: 1.232

Sample information

Sample name: Human metapneumovirus matrix protein M bound to DOPC
Ligand: 1,2-Dioleoyl-sn-glycero-3-phosphocholine
Protein: human metapneumovirus matrix protein

Validation information

Experimental information

 

Image processing

Software: Burnham-Brandeis, Helical, Package
Reconstruction protocol: Helical reconstruction
CTF correction: Each micrograph
Details: Final map was filtered to 28 angstrom resolution The reconstruction was calculated using Burnham-Brandeis Helical Package

Imaging

Session
Microscope model: FEI TECNAI F30
Electron source: FIELD EMISSION GUN
Nominal magnification: 39000.0
Defocus max (nm): 820.00
Defocus min (nm): 720.00
Holder model: SIDE ENTRY, EUCENTRIC
Acquisition
Number of images: 1
Sampling interval (μm): 15.0

Specimen

Preparation
Specimen state: filament
Specimen concentration (mg/mL): 0.20
Buffer: 20 mM Tris, 650 mM NaCl, 1M NDSB-201
Staining procedure: Grids with adsorbed protein-lipid mixtures floated on 2% w/v uranyl acetate for 30 seconds
Specimen support: 300 mesh copper grid with formvar carbon film
Aggregation parameters (helical parameters)
Phi increment (degrees): 56.5
Azimuthal (Z) increment (Å): 5.1600000000000001
Vitrification
Apparatus: NONE
Cryogen: NONE