5TCR
Atomic model of the Salmonella SPI-1 type III secretion injectisome basal body proteins InvG, PrgH, and PrgK
ELECTRON MICROSCOPY
| Sample |
|---|
| Type III injectisome basal body |
| 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 | 67800 |
| Reported Resolution (Å) | 6.3 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | Limited frequency refinement in Frealign |
| Refinement Type | |
| Symmetry Type | POINT |
| Point Symmetry | C1 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | FLEXIBLE FIT | ||||
| Refinement Target | |||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | Initial fitting was carried out with Chimera, followed by rebuilding and refinement in Rosetta, Phenix, and Coot. | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K2 QUANTUM (4k x 4k) | ||||||||
| Electron Dose (electrons/Å**2) | 1.3 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | |
| Temperature (Kelvin) | |
| Microscope Model | FEI TITAN KRIOS |
| Minimum Defocus (nm) | 1500 |
| Maximum Defocus (nm) | 3000 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | 0.01 |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | FEI TITAN KRIOS AUTOGRID HOLDER |
| Nominal Magnification | 64000 |
| Calibrated Magnification | 29240 |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 300 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| PARTICLE SELECTION | EMAN2 | |
| PARTICLE SELECTION | RELION | |
| IMAGE ACQUISITION | SerialEM | |
| CTF CORRECTION | CTFFIND | |
| INITIAL EULER ASSIGNMENT | RELION | |
| INITIAL EULER ASSIGNMENT | FREALIGN | |
| FINAL EULER ASSIGNMENT | FREALIGN | |
| RECONSTRUCTION | FREALIGN | |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| PHASE FLIPPING AND AMPLITUDE CORRECTION | 263900 | |||
