7AD3
Class D GPCR Ste2 dimer coupled to two G proteins
ELECTRON MICROSCOPY
Refinement
| RMS Deviations | |
|---|---|
| Key | Refinement Restraint Deviation |
| f_dihedral_angle_d | 8.558 |
| f_angle_d | 0.894 |
| f_chiral_restr | 0.053 |
| f_plane_restr | 0.006 |
| f_bond_d | 0.005 |
| Sample |
|---|
| Ste2 dimer coupled to two G proteins |
| Sample Components |
| Ste2 dimer coupled to two G proteins |
| Alpha-factor mating pheromone |
| Guanine nucleotide-binding protein alpha-1 subunit,Guanine nucleotide-binding protein alpha-1 subunit |
| 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 | 96611 |
| Reported Resolution (Å) | 3.5 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | |
| Refinement Type | |
| Symmetry Type | POINT |
| Number of Particles | 16418 |
| Reported Resolution (Å) | 3.3 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | 16418 particles from this dataset contributed to the final 131274 particles that provided map 2 at 3.3 angstrom global resolution |
| Refinement Type | |
| Symmetry Type | POINT |
| Number of Particles | 4556 |
| Reported Resolution (Å) | 3.3 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | 4556 particles from this dataset contributed to the final 131274 particles that provided map 2 at 3.3 angstrom global resolution |
| Refinement Type | |
| Symmetry Type | POINT |
| Number of Particles | 110300 |
| Reported Resolution (Å) | 3.3 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | 110300 particles from this dataset contributed to the final 131274 particles that provided map 2 at 3.3 angstrom global resolution |
| Refinement Type | |
| Symmetry Type | POINT |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 (6G79, 6G79, 6G79) | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | |||||
| Refinement Target | Correlation coefficient | ||||
| Overall B Value | 112 | ||||
| Fitting Procedure | |||||
| Details | Manual building was performed in Coot iterated with real space refinement in PHENIX. | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K2 QUANTUM (4k x 4k) | GATAN K2 QUANTUM (4k x 4k) | GATAN K2 QUANTUM (4k x 4k) | ||||||
| Electron Dose (electrons/Å**2) | 45 | 48 | 40 | ||||||
| Imaging Experiment | 1 | 2 | 3 |
|---|---|---|---|
| Date of Experiment | |||
| Temperature (Kelvin) | |||
| Microscope Model | FEI TITAN KRIOS | FEI TITAN KRIOS | FEI TITAN KRIOS |
| Minimum Defocus (nm) | 900 | 900 | 900 |
| Maximum Defocus (nm) | 2700 | 2700 | 2700 |
| Minimum Tilt Angle (degrees) | |||
| Maximum Tilt Angle (degrees) | |||
| Nominal CS | 2.7 | 2.7 | 2.7 |
| Imaging Mode | BRIGHT FIELD | BRIGHT FIELD | BRIGHT FIELD |
| Specimen Holder Model | FEI TITAN KRIOS AUTOGRID HOLDER | FEI TITAN KRIOS AUTOGRID HOLDER | FEI TITAN KRIOS AUTOGRID HOLDER |
| Nominal Magnification | 130000 | 105000 | 105000 |
| Calibrated Magnification | |||
| Source | FIELD EMISSION GUN | FIELD EMISSION GUN | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 300 | 300 | 300 |
| Imaging Details | eBIC Krios1 | LMB Krios1 | LMB Krios 2 |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| MODEL FITTING | Coot | 0.9 pre-EL |
| INITIAL EULER ASSIGNMENT | RELION | 3.1 |
| FINAL EULER ASSIGNMENT | RELION | 3.1 |
| RECONSTRUCTION | RELION | 3.1 |
| INITIAL EULER ASSIGNMENT | RELION | 3.1 |
| FINAL EULER ASSIGNMENT | RELION | 3.1 |
| RECONSTRUCTION | RELION | 3.1 |
| INITIAL EULER ASSIGNMENT | RELION | 3.1 |
| FINAL EULER ASSIGNMENT | RELION | 3.1 |
| RECONSTRUCTION | RELION | |
| INITIAL EULER ASSIGNMENT | RELION | 3.1 |
| FINAL EULER ASSIGNMENT | RELION | 3.1 |
| MODEL REFINEMENT | PHENIX | 1.17.1-3660 |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| NONE | 2193519 | |||
| NONE | 485568 | |||
| NONE | 781447 | |||
| NONE | 2193519 | |||
