EMD-2328 Experiments and Validation

Structure of the bacterial V-ATPase from Thermus thermophilius

Electron crystallography
Overview of EMD-2328
Sample name: Bacterial V-type ATPase
Organism: Thermus thermophilus

Map parameters

Recommended contour level: 0.40 (author)
Number of grid points: 31 × 55 × 73
Voxel size: 4.30 × 4.40 × 4.17 Å
Minimum density: -3.489
Maximum density: 6.748
Average density: -0.006
Standard deviation: 0.987

Sample information

Sample name: Bacterial V-type ATPase
Protein: V-ATPase

Validation information

Experimental information

 

Image processing

Software: MRC

Imaging

Session
Microscope model: FEI TECNAI F30
Electron source: FIELD EMISSION GUN
Nominal magnification: 40000.0
Defocus max (nm): 2,270.00
Defocus min (nm): 1,400.00
Tilt max (degrees): 55.00
Tilt min (degrees): -55.00
Holder model: SIDE ENTRY, EUCENTRIC
Acquisition
Number of images: 212
Sampling interval (μm): 15.0
Details: Every image was recorded by 2Kx2K CCD camera (Gatan)

Specimen

Preparation
Specimen state: twoDArray
Specimen concentration (mg/mL): 1.00
Staining procedure: A 2.5 ul sample was transferred to a glow-discharged carbon coated copper grid. After 1 min, the drop was blotted with a slice of filter paper, and then the grid was washed with 2.5 ul water to avoid the precipitation caused by mixing phosphate buffer with uranyl acetate. Subsequently, the sample was stained with 2.5 ul of 2% uranyl acetate and air-dried.
Specimen support: a glow-discharged carbon coated copper grid
Crystal grow: Crystals grown by dialysis
Aggregation parameters (crystal parameters)
Plane group: P 1
Unit cell length A (Å): 232.0
Unit cell length B (Å): 132.0
Unit cell angle gamma (degrees): 90.0
Vitrification
Apparatus: NONE
Cryogen: NONE