Atomic model of a microtubule seam based on a cryo-EM reconstruction of the EB3-bound microtubule (merged dataset containing tubulin bound to GTPgammaS, GMPCPP, and GDP)
ELECTRON MICROSCOPY
Refinement
| RMS Deviations | |
|---|---|
| Key | Refinement Restraint Deviation |
| r_dihedral_angle_2_deg | 37.202 |
| r_dihedral_angle_3_deg | 16.948 |
| r_dihedral_angle_4_deg | 13.25 |
| r_dihedral_angle_1_deg | 8.85 |
| r_mcangle_it | 5.509 |
| r_mcbond_it | 3.032 |
| r_mcbond_other | 3.032 |
| r_angle_refined_deg | 1.379 |
| r_angle_other_deg | 0.947 |
| r_chiral_restr | 0.081 |
| Sample |
|---|
| EB3-bound microtubule (merged dataset containing tubulin bound to GTPgammaS, GMPCPP and GDP) |
| 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 | 79702 |
| Reported Resolution (Å) | 3.9 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | asymmetric (C1) reconstruction |
| Refinement Type | |
| Symmetry Type | HELICAL |
| Axial Symmetry | C1 |
| Axial Rise | 9.44 |
| Angular Rotation | 27.71 |
| 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 | ||||
