EMD-22009 Experiments and Validation

Structure of human TRPA1 in complex with agonist GNE551

Single particle reconstruction
Overview of EMD-22009
Sample name: TRPA1 bound by agonist GNE551
Organism: Homo sapiens
Fitted atomic model: 6x2j

Map parameters

Recommended contour level: 0.20 (author)
Number of grid points: 324 × 324 × 324
Voxel size: 1.00 × 1.00 × 1.00 Å
Minimum density: -1.848
Maximum density: 2.837
Average density: 0.00
Standard deviation: 0.048

Sample information

Sample name: TRPA1 bound by agonist GNE551
Ligand: 5-amino-1-[(4-bromo-2-fluorophenyl)methyl]-N-(2,5-dimethoxyphenyl)-1H-1,2,3-triazole-4-carboxamide
Proteins: TRPA1 bound by agonist GNE551, Transient receptor potential cation channel subfamily A member 1

Validation information

Experimental information

 

Image processing

Applied symmetry: C4
Software: cisTEM
Number of particles: 58837
Reconstruction protocol: FOURIER SPACE
Details: Data at spatial frequencies higher than 1/4.0 A-1 were not used during any part of the refinement. The final map was density-modified using Phenix.

Fitting

Initial atomic model: 6PQQ
Refinement space: REAL

Imaging

Session
Microscope model: FEI TITAN KRIOS
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 41.799999999999997
Energy filter: GIF Bioquantum
Nominal CS (mm): 2.70
Nominal magnification: 165000.0
Holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Acquisition
Number of images: 11063
Sampling interval (μm): 5.0

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 0.33
Specimen support: Solarus plasma cleaner
Vitrification
Apparatus: FEI VITROBOT MARK IV
Cryogen: ETHANE
Humidity: 100
Details: Triple blot. Put blot in vitroblot Apply 3.5ul to grid, wait 30sec, blot manually Apply 3.5ul to grid, wait 30sec, blot manually, apply final 3.5ul, final blot by vitrobot (3.5s).