EMD-2299 Experiments and Validation

Negative stain EM structure of the PTC3 holotoxin complex (TcdA1, TcdB2, TccC3) in prepore state

Single particle reconstruction
Overview of EMD-2299
Sample name: PTC3 (TcdA1, TcdB2, TccC3)
Organism: Photorhabdus luminescens
Related EM entries by publication: EMD-2301, EMD-2300, EMD-2298, EMD-2297

Map parameters

Minimum density: -0.059
Maximum density: 0.097
Average density: 0.001
Standard deviation: 0.007
Recommended contour level: 0.021 (author)

Sample information

Sample name: PTC3 (TcdA1, TcdB2, TccC3)
Proteins: TcdA1, TccC3, TcdB2

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Sparx
Number of particles: 3022
Reconstruction protocol: Projection Matching

Imaging

Session
Electron source: LAB6
Nominal CS (mm): 3.40
Nominal magnification: 50000.0
Defocus max (nm): 2.00
Defocus min (nm): 1.20
Holder model: JEOL
Acquisition
Number of images: 84

Specimen

Preparation
Specimen state: particle
Buffer: 50 mM MES, 100 mM NaCl, 0.05% Tween-20, 5% glycerol
Staining procedure: Grids with adsorbed Protein floated on 0.07% Uranyl Formate for 45 sec.
Specimen support: glow discharged 400 mesh copper grids with thin carbon support
Vitrification
Apparatus: NONE
Cryogen: NONE