EMD-2271 Experiments and Validation

Cryo EM map of the Gaalphaq-PLCbeta3 complex.

Single particle reconstruction
Overview of EMD-2271
Sample name: Cryo EM reconstruction of the Galphaq-PLCbeta3 complex.
Organisms: Mus musculus, Homo sapiens
Related EM entry by publication: EMD-5561

Map parameters

Minimum density: -1.97
Maximum density: 6.23
Average density: 0
Standard deviation: 0.35
Recommended contour level: 0.84 (author)

Sample information

Sample name: Cryo EM reconstruction of the Galphaq-PLCbeta3 complex.
Proteins: G alpha q, phospholipase C beta 3

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: EMAN1
Number of particles: 12673
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
Nominal magnification: 50000.0
Defocus max (nm): 3,500
Defocus min (nm): 1,500
Holder model: OTHER
Acquisition
Number of images: 350

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1
Buffer: 20 mM HEPES pH 8, 200 mM NaCl, 2 mM DTT, 0.9 mM CaCl2, 50 microM GDP, 30 microM 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 for 2 seconds at 3 microliter sample.
Details: Vitrification carried out in liquid nitrogen atmosphere.