ELECTRON MICROSCOPY
| Sample |
|---|
| THERMUS THERMOPHILUS 30S ribosomal subunit complexed with Era |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | PARTICLE |
| Vitrification Instrument | |
| Cryogen Name | ETHANE |
| Sample Vitrification Details | Rapid-freezing in liquid ethane |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | SINGLE PARTICLE |
| Number of Particles | |
| Reported Resolution (Å) | 13.5 |
| Resolution Method | |
| Other Details | projection matching using spider package. The coordinates for only the alpha carbons in protein and phosphoruses in nucleic acid are present in the s ... |
| Refinement Type | |
| Symmetry Type | POINT |
| Point Symmetry | C1 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 (1FJF, 1EGA) | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | RIGID BODY FIT | ||||
| Refinement Target | X-ray coordinates of the 30S ribosomal subunit and era were fitted into the 13.5 angstroms resolution CRYO-EM map of the T. Thermophilus 30S subunit-era complex. The atomic structure of era was fitted as 3 rigid bodies, N-terminal domain, C-terminal domain and C-terminal helix within the C-terminal domain. the resultant era structure was then energy minimized. The X-ray coordinates of T. Thermophilus 30S subunit was fitted as 4 rigid bodies, head, body, platform and 16S rRNA 3' minor domains. Only the proteins and segments of RNA helices of the 30S subunit that contact era in the era-30S complex are included here. | ||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | METHOD--Cross-correlation coefficient based manual fitting in O REFINEMENT PROTOCOL--MULTIPLE RIGID BODY | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | KODAK SO-163 FILM | ||||||||
| Electron Dose (electrons/Å**2) | 20 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | 2003-03-25 |
| Temperature (Kelvin) | 93 |
| Microscope Model | FEI TECNAI F20 |
| Minimum Defocus (nm) | 1180 |
| Maximum Defocus (nm) | 3940 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | 2 |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | |
| Nominal Magnification | 50000 |
| Calibrated Magnification | 49696 |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 200 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| MODEL FITTING | O | |
| RECONSTRUCTION | SPIDER | |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| CTF correction of 3D-MAPS by wiener filtration | ||||
