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).