ELECTRON MICROSCOPY

Sample

Local refinement of the N-domain of full-length, soluble, monomeric somatic angiotensin I-converting enzyme

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details Diluted protein (in buffer containing zinc chloride and sodium chloride) was incubated on ice for 30 minutes after which it was applied to the grid, incubated for 30 seconds, and blotted for 3 seconds before plunging
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 43774
Reported Resolution (Å) 3.72
Resolution Method FSC 0.143 CUT-OFF
Other Details Local refinement with a mask around the N-domain
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol FLEXIBLE FIT
Refinement Target Correlation coefficient
Overall B Value ?
Fitting Procedure ?
Details ?
Data Acquisition
Detector Type GATAN K3 (6k x 4k)
Electron Dose (electrons/Å2) 43
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model FEI TITAN KRIOS
Minimum Defocus (nm) 1800
Maximum Defocus (nm) 3000
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS 2.7
Imaging Mode BRIGHT FIELD
Specimen Holder Model FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification 81000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION Topaz 0.2.5
IMAGE ACQUISITION EPU ?
CTF CORRECTION cryoSPARC 3.2
MODEL FITTING ISOLDE 1.2.0
MODEL REFINEMENT PHENIX 1.19.2
INITIAL EULER ASSIGNMENT cryoSPARC 3.2
FINAL EULER ASSIGNMENT cryoSPARC 3.2
CLASSIFICATION RELION 3.1
RECONSTRUCTION cryoSPARC 3.2
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION cryoSPARC patch-based CTF estimation
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