EMD-1980 Experiments and Validation

CryoEM map of ParM filament at 7.2 Angstrom resolution

Helical reconstruction
Overview of EMD-1980
Sample name: ParM filament
Organism: Escherichia coli
Fitted atomic model: 4a6j

Map parameters

Recommended contour level: 0.631 (author)
Number of grid points: 100 × 100 × 100
Voxel size: 1.64 × 1.64 × 1.64 Å
Minimum density: -1.246
Maximum density: 1.864
Average density: 0.003
Standard deviation: 0.209

Sample information

Sample name: ParM filament
Protein: Plasmid segregation protein parM

Validation information

Experimental information

 

Image processing

Reconstruction protocol: Projection Matching
CTF correction: CTFFIND3 Each particle

Fitting

Initial atomic model: 4A62
Used chain: 4A62_A
Fitting software: Chimera
Fitting protocol: rigid body
Refinement space: REAL
Details: Protocol: Rigid Body

Imaging

Session
Microscope model: JEOL 3200FSC
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Energy filter: Omega filter
Energy window (eV): 0.00
Nominal CS (mm): 1.60
Nominal magnification: 50000.0
Holder model: JEOL
Acquisition
Number of images: 207
Sampling interval (μm): 15.0

Specimen

Preparation
Specimen state: filament
Buffer: 30 mM Tris-HCl, 25 mM KCl, 2 mM MgCl2, 1 mM DTT, pH 7.5
Specimen support: Quantifoil holey carbon molybdenum grid (R0.6/1.0, Quantifoil Micro Tools GmbH, Jena, Germany)
Aggregation parameters (helical parameters)
Handedness: RIGHT HANDED
Phi increment (degrees): 164.97999999999999
Azimuthal (Z) increment (Å): 23.620999999999999
Vitrification
Apparatus: FEI VITROBOT
Cryogen: ETHANE
Humidity: 100
Protocol: Blot for 3.5 seconds before plunging
Details: Vitrification instrument: Vitrobot