2BCW
Coordinates of the N-terminal domain of ribosomal protein L11,C-terminal domain of ribosomal protein L7/L12 and a portion of the G' domain of elongation factor G, as fitted into cryo-em map of an Escherichia coli 70S*EF-G*GDP*fusidic acid complex
ELECTRON MICROSCOPY
| Sample |
|---|
| 70S*EF-G*GDP*FUSIDIC ACID COMPLEX |
| Sample Components |
| 50S ribosomal protein L11 |
| 50S ribosomal protein L7/L12 |
| Elongation factor G |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | PARTICLE |
| Vitrification Instrument | |
| Cryogen Name | |
| Sample Vitrification Details | RAPID-FREEZING IN LIQUID ETHANE |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | SINGLE PARTICLE |
| Number of Particles | |
| Reported Resolution (Å) | 11.2 |
| Resolution Method | |
| Other Details | PROJECTION MATCHING USING SPIDER PACKAGE |
| Refinement Type | |
| Symmetry Type | POINT |
| Point Symmetry | C1 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 (1MMS, 1CTF, 1ELO) | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | FLEXIBLE FIT | ||||
| Refinement Target | The X-ray crystallographic structure of the EF-G individual domains were fitted into the cryo-EM map of the 70S*EF-G-UG*GDP*Fusidic acid complex using O. The G' domain within domain I was separately fitted, using a combination of manual rigid-body docking and flexible docking approaches, and taking into consideration both the cryo-EM envelope and the positional constraints imposed by the UG density. X-ray crystallographic structures of the N-terminal domain of protein L11 and C-terminal domain of protein L7/L12 were fitted as rigid bodies. | ||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | METHOD--A COMBINATION OF MANUAL RIGID-BODY DOCKING AND FLEXIBLE DOCKING REFINEMENT PROTOCOL--A COMBINATION OF MANUAL RIGID-BODY DOCKING AND FLEXIBLE ... | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | KODAK SO-163 FILM | ||||||||
| Electron Dose (electrons/Å**2) | 20 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | 2004-11-09 |
| Temperature (Kelvin) | 93 |
| Microscope Model | FEI TECNAI F20 |
| Minimum Defocus (nm) | -0.7 |
| Maximum Defocus (nm) | -3.5 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | 2 |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | |
| Nominal Magnification | 50000 |
| Calibrated Magnification | 50760 |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 200 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| MODEL FITTING | OTHER | |
| 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 | ||||
