EMD-5679 Experiments and Validation

Electron Microscopy of the Aquaporin-0/Calmodulin Complex

Single particle reconstruction
Overview of EMD-5679
Sample name: Aquaporin-0 bound to Calmodulin
Organisms: Ovis aries, Homo sapiens
Fitted atomic model: 3j41

Map parameters

Recommended contour level: 4.96 (author)
Number of grid points: 66 × 66 × 66
Voxel size: 3.98 × 3.98 × 3.98 Å
Minimum density: -3.114
Maximum density: 8.374
Average density: -0.00
Standard deviation: 1.00

Sample information

Sample name: Aquaporin-0 bound to Calmodulin
Proteins: Aquaporin-0, Calmodulin

Validation information

Experimental information


Image processing

Applied symmetry: C2
Number of particles: 11720
Reconstruction protocol: Random Conical Tilt
CTF correction: CTF-TILT, each micrograph
Details: Final Map with C2 Symmetry and Filtered to 25 Angstrom Particles were selected from a tilted pair dataset at 0 and 50 degree tilt using SPIDER. An initial reconstruction was generated using random conical tilt methods in SPIDER and refined in FREALIGN


Initial atomic model: 2B6P
Used chain: A
Fitting software: Chimera
Fitting protocol: rigid body
Target criteria: cross-correlation
Refinement space: REAL


Microscope model: FEI TECNAI 12
Electron source: LAB6
Electron dose (e/A**2): 15.0
Nominal CS (mm): 2.00
Nominal magnification: 52000.0
Defocus max (nm): 2,000.00
Defocus min (nm): 1,000.00
Tilt max (degrees): 50.00
Tilt min (degrees): 0.00
Holder model: OTHER
Number of images: 200
Scanner model: NIKON SUPER COOLSCAN 9000
Sampling interval (μm): 6.3499999999999996


Specimen state: particle
Specimen concentration (mg/mL): 0.02
Buffer: 25mM HEPES, 5mM CaCl2, 0.3% decylmaltoside
Staining procedure: 0.75% uranyl formate
Specimen support: 400 mesh carbon coated grid (Ted Pella)
Apparatus: NONE
Cryogen: NONE