Cryo-EM structure of GTPgammaS-microtubule co-polymerized with EB3 (merged dataset with and without kinesin bound)
ELECTRON MICROSCOPY
Refinement
| RMS Deviations | |
|---|---|
| Key | Refinement Restraint Deviation |
| r_dihedral_angle_2_deg | 35.041 |
| r_dihedral_angle_3_deg | 14.078 |
| r_dihedral_angle_1_deg | 11.414 |
| r_dihedral_angle_4_deg | 10.893 |
| r_mcangle_it | 1.888 |
| r_angle_refined_deg | 1.331 |
| r_mcbond_it | 1.006 |
| r_mcbond_other | 1.006 |
| r_angle_other_deg | 0.931 |
| r_chiral_restr | 0.077 |
| Sample |
|---|
| EB3- and kinesin-bound GTPgammaS microtubule |
| Sample Components |
| Alpha tubulin |
| Beta tubulin |
| EB3 |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | FILAMENT |
| Vitrification Instrument | FEI VITROBOT MARK II |
| Cryogen Name | ETHANE |
| Sample Vitrification Details | Blot once for 4 seconds before plunging into liquid ethane (FEI VITROBOT MARK II). |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | HELICAL |
| Number of Particles | |
| Reported Resolution (Å) | 3.3 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | IHRSR algorithm with microtubule-specific pseudo-helical symmetry applied |
| Refinement Type | |
| Symmetry Type | HELICAL |
| Axial Symmetry | C1 |
| Axial Rise | 9.45 |
| Angular Rotation | 27.72 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 | ||||
| Refinement Space | |||||
| Refinement Protocol | |||||
| Refinement Target | |||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | |||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K2 (4k x 4k) | ||||||||
| Electron Dose (electrons/Å**2) | 27.6 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | 2013-05-16 |
| Temperature (Kelvin) | 90 |
| Microscope Model | FEI TITAN |
| Minimum Defocus (nm) | 1000 |
| Maximum Defocus (nm) | 3500 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | 2.7 |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | GATAN LIQUID NITROGEN |
| Nominal Magnification | 27500 |
| Calibrated Magnification | 27500 |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 300 |
| Imaging Details |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| RECONSTRUCTION | EMAN | 1 |
| RECONSTRUCTION | FREALIGN | |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| CTFFIND4, each particle | ||||
