EMD-2102 Experiments and Validation

Structure of a stalled transfer intermediate of Sm proteins from the assembly chaperone pICln to the SMN-complex

Single particle reconstruction
Overview of EMD-2102
Sample name: 8S complex: a stalled Sm protein transfer intermediate from the assembly chaperone pICln to the SMN-complex
Organism: Homo sapiens

Map parameters

Minimum density: -0.24
Maximum density: 1.33
Average density: 0.01
Standard deviation: 0.07
Recommended contour level: 0.02 (author)

Sample information

Sample name: 8S complex: a stalled Sm protein transfer intermediate from the assembly chaperone pICln to the SMN-complex
Protein: 8S complex

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Imagic
Number of particles: 10000
Reconstruction protocol: exact filter backprojection
CTF correction: Each particle

Fitting

Initial atomic model: 4F77
Fitting software: Amira
Fitting protocol: rigid body
Refinement space: REAL
Details: Protocol: Rigid Body

Imaging

Session
Microscope model: FEI/PHILIPS CM200FEG
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Nominal CS (mm): 2
Holder model: SIDE ENTRY, EUCENTRIC
Acquisition
Number of images: 48

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 10
Buffer: 20 mM HEPES, 150 mM NaCl, 5 mM DTT
Staining procedure: Grids were floated on 2 % Uranyl Formate solution for 1 minute.
Specimen support: Custom made holey grid with thin carbon support.
Vitrification
Apparatus: NONE
Cryogen: NONE
Details: negative stain