EMD-20762 Experiments and Validation

Katanin hexamer in the ring conformation in complex with substrate

Single particle reconstruction
Overview of EMD-20762
Sample name: Hexameric complex of C.elegans katanin bound to polyglutamate peptide
Organisms: Caenorhabditis elegans, synthetic construct
Fitted atomic model: 6uge

Map parameters

Recommended contour level: 5.74 (author)
Number of grid points: 224 × 224 × 224
Voxel size: 1.08 × 1.08 × 1.08 Å
Minimum density: -17.77
Maximum density: 30.531
Average density: 0.00
Standard deviation: 1.00

Sample information

Sample name: Hexameric complex of C.elegans katanin bound to polyglutamate peptide
Ligands: ADENOSINE-5'-TRIPHOSPHATE, MAGNESIUM ION
Proteins: Hexameric complex of C.elegans katanin bound to polyglutamate peptide, Meiotic spindle formation protein mei-1, Polyglutamate peptide

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: RELION
Number of particles: 108700
Reconstruction protocol: FOURIER SPACE

Fitting

Initial atomic model: 5WC0
Fitting protocol: flexible
Target criteria: Correlation coefficient
Refinement space: REAL

Imaging

Session
Microscope model: FEI TITAN KRIOS
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 73.0
Energy filter: GIF Quantum LS
Nominal CS (mm): 2.70
Nominal magnification: 130000.0
Defocus max (nm): 2,800.00
Defocus min (nm): 1,000.00
Holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Acquisition
Number of images: 5911

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1.00
Vitrification
Apparatus: LEICA EM GP
Cryogen: ETHANE
Humidity: 90
Details: Load 5 ul sample, wait 10 sec, blot 4.5 sec.