Hybrid model of a complex of BREX proteins BrxB and PglZ from Salmonella typhimurium
ELECTRON MICROSCOPY
| Sample |
|---|
| Complex of BREX proteins BrxB / PglZ from Salmonella Typhimurium str. D23580 |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | PARTICLE |
| Vitrification Instrument | FEI VITROBOT MARK IV |
| Cryogen Name | ETHANE |
| Sample Vitrification Details | |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | SINGLE PARTICLE |
| Number of Particles | 123964 |
| Reported Resolution (Å) | 4.45 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | |
| Refinement Type | |
| Symmetry Type | POINT |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | RIGID BODY FIT | ||||
| Refinement Target | |||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | AlphaFold model, split into domains at "hinge" points, was rigid fit into the cryo-EM map. The following domains were used: (1) BrxB + 1-96 PglZ, (2) ... | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K3 (6k x 4k) | ||||||||
| Electron Dose (electrons/Å**2) | 50 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | |
| Temperature (Kelvin) | |
| Microscope Model | TFS GLACIOS |
| Minimum Defocus (nm) | 800 |
| Maximum Defocus (nm) | 2000 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | 2.7 |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | |
| Nominal Magnification | 36000 |
| Calibrated Magnification | |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 200 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| PARTICLE SELECTION | cryoSPARC | 4.2.1 |
| PARTICLE SELECTION | cryoSPARC | 4.2.1 |
| IMAGE ACQUISITION | SerialEM | |
| CTF CORRECTION | cryoSPARC | 4.2.1 |
| MODEL FITTING | UCSF ChimeraX | 1.9 |
| MODEL FITTING | ISOLDE | 1.6 |
| INITIAL EULER ASSIGNMENT | cryoSPARC | 4.2.1 |
| FINAL EULER ASSIGNMENT | cryoSPARC | 4.2.1 |
| CLASSIFICATION | cryoSPARC | 4.2.1 |
| RECONSTRUCTION | cryoSPARC | 4.2.1 |
| MODEL REFINEMENT | PHENIX | |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| PHASE FLIPPING AND AMPLITUDE CORRECTION | 15593169 | 10,876,932 from blob picking on dataset 1 4,716,237 from template picking on dataset 2 | ||
