EMD-2069 Experiments and Validation

Octameric structure of IMPDH from Pseudomonas aeruginosa in its APO form

Single particle reconstruction
Overview of EMD-2069
Sample name: IMPDH apo filtered at 15.5 A resolution from Pseudomonas aeruginosa
Organism: Pseudomonas aeruginosa
Related EM entry by publication: EMD-2070

Map parameters

Recommended contour level: 45.00 (author)
Number of grid points: 150 × 150 × 150
Voxel size: 2.30 × 2.30 × 2.30 Å
Minimum density: 0.00
Maximum density: 100.00
Average density: 31.148
Standard deviation: 5.581

Sample information

Sample name: IMPDH apo filtered at 15.5 A resolution from Pseudomonas aeruginosa
Protein: Inosine-5'-monophosphate dehydrogenase

Validation information

Experimental information

 

Image processing

Applied symmetry: D4
Software: IMAGIC, FREALIGN
Number of particles: 10625
CTF correction: Each particle

Imaging

Session
Microscope model: JEOL 2200FS
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 18.0
Energy filter: JEOL
Energy window (eV): 0.00
Nominal CS (mm): 2.00
Nominal magnification: 100000.0
Defocus max (nm): 2,500.00
Defocus min (nm): 1,400.00
Holder model: GATAN LIQUID NITROGEN
Details: Weak beam illumination
Acquisition
Number of images: 74
Details: Images recorded with the 4K X 4K slow-scan CCD camera (Gatan inc.), which explains why the nominal magnification is 100000 and 130137 when calibrated on the camera.

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1.00
Buffer: 20 mM potassium phosphate pH 8 and 0.1 M KCl
Specimen support: Quantifoil R2.2, glow discharged
Vitrification
Apparatus: GATAN CRYOPLUNGE 3
Cryogen: ETHANE
Humidity: 98
Protocol: Blot for 1 second