EMD-5561 Experiments and Validation

Cryo EM map of the Gaalphaq-PLCbeta3 complex.

Single particle reconstruction
Overview of EMD-5561
Sample name: Reconstruction of the PLCbeta3 distal CTD in complex with the N-terminal helix of Galphaq
Organisms: Mus musculus, Homo sapiens
Related EM entry by publication: EMD-2271

Map parameters

Recommended contour level: 2.50 (author)
Number of grid points: 64 × 64 × 64
Voxel size: 4.48 × 4.48 × 4.48 Å
Minimum density: -6.214
Maximum density: 18.742
Average density: 0.001
Standard deviation: 1.266

Sample information

Sample name: Reconstruction of the PLCbeta3 distal CTD in complex with the N-terminal helix of Galphaq
Proteins: G alpha q, phospholipase C beta 3

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: EMAN1
Number of particles: 12692
Reconstruction protocol: cross-common lines
CTF correction: each particle

Fitting

Initial atomic model: 4GNK
Refinement space: REAL

Imaging

Session
Microscope model: FEI TECNAI F20
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Nominal CS (mm): 2.00
Nominal magnification: 50000.0
Defocus max (nm): 3,500.00
Defocus min (nm): 1,500.00
Holder model: OTHER
Acquisition
Number of images: 350

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1.00
Buffer: 20 mM HEPES, pH 8, 200 mM NaCl, 2 mM DTT, 0.9 mM CaCl2, 50 uM GDP, 30 uM AlCl3, 10 mM NaF, 5 mM MgCl2
Specimen support: glow-discharged Quantifoil R2/200 mesh grid
Vitrification
Apparatus: FEI VITROBOT MARK IV
Cryogen: ETHANE
Humidity: 100
Protocol: Blot 3 microliter sample for 2 seconds.
Details: Vitrification carried out in liquid nitrogen atmosphere.