EMD-2298 Experiments and Validation

Structure of TcdA1 pore (TcdA1 reconstituted in liposomes)

Single particle reconstruction
Overview of EMD-2298
Sample name: TcdA1
Organism: Photorhabdus luminescens
Related EM entries by publication: EMD-2297, EMD-2299, EMD-2300, EMD-2301

Map parameters

Recommended contour level: 0.012 (emdb)
Number of grid points: 128 × 128 × 128
Voxel size: 3.65 × 3.65 × 3.65 Å
Minimum density: -0.035
Maximum density: 0.075
Average density: 0.001
Standard deviation: 0.006

Sample information

Sample name: TcdA1
Protein: TcdA1

Validation information

Experimental information

 

Image processing

Applied symmetry: C5
Software: Sparx
Number of particles: 5549
Reconstruction protocol: Projection matching
CTF correction: Each Particle

Imaging

Session
Microscope model: JEOL 3200FSC
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Energy filter: in-column Omega filter
Energy window (eV): 0.00
Nominal CS (mm): 4.10
Nominal magnification: 40000.0
Defocus max (nm): 5.00
Defocus min (nm): 2.90
Holder model: JEOL 3200FSC CRYOHOLDER
Acquisition
Number of images: 145

Specimen

Preparation
Specimen state: particle
Buffer: 50 mM CAPS pH 11, 100 mM NaCl, 5 % glycerol
Specimen support: C-Flat 1.2/1.3-4C copper 400 mesh, with additional thin carbon support
Vitrification
Apparatus: GATAN CRYOPLUNGE 3
Cryogen: ETHANE
Humidity: 90
Protocol: Blot for 1 sec before plunging