EMD-5498 Experiments and Validation

Structure of ABCA4, the photoreceptor-specific ATP-binding cassette transporter, in complex with AMPPNP

Single particle reconstruction
Overview of EMD-5498
Sample name: ABCA4 purified from bovine rod outer segments and supplemented with AMPPNP, a non-hydrolyzable ATP analogue
Organism: Bos taurus
Related EM entry by publication: EMD-5497

Map parameters

Recommended contour level: 0.036 (author)
Number of grid points: 144 × 144 × 144
Voxel size: 2.13 × 2.13 × 2.13 Å
Minimum density: -0.073
Maximum density: 0.117
Average density: -0.002
Standard deviation: 0.01

Sample information

Sample name: ABCA4 purified from bovine rod outer segments and supplemented with AMPPNP, a non-hydrolyzable ATP analogue
Protein: ABCA4

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Spider
Number of particles: 38090
Reconstruction protocol: reference-based projection alignment
CTF correction: Each defocus group

Imaging

Session
Microscope model: FEI TECNAI F20
Electron source: FIELD EMISSION GUN
Nominal magnification: 50000.0
Defocus max (nm): 1,500.00
Defocus min (nm): 200.00
Tilt max (degrees): 45.00
Tilt min (degrees): 0.00
Holder model: OTHER
Acquisition
Number of images: 200

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 0.10
Buffer: 20 mM BTP, pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, 1 mM AMPPNP
Staining procedure: Grids with adsorbed protein were stained with 1% w/v uranyl acetate for 10 seconds
Specimen support: Glow discharged Cu/Pd 400 mesh grids with thin carbon support.
Vitrification
Apparatus: NONE
Cryogen: NONE