Structure of human HCN4 hyperpolarization-activated cyclic nucleotide-gated ion channel
ELECTRON MICROSCOPY
| Sample |
|---|
| Potassium/Sodium hyperpolarization-activated cyclic nucleotide-gated ion channel 4 |
| Specimen Preparation | |
|---|---|
| Sample Aggregation State | PARTICLE |
| Vitrification Instrument | FEI VITROBOT MARK IV |
| Cryogen Name | ETHANE |
| Sample Vitrification Details | blotted for 5.5s before plunge |
| 3D Reconstruction | |
|---|---|
| Reconstruction Method | SINGLE PARTICLE |
| Number of Particles | 25764 |
| Reported Resolution (Å) | 3.4 |
| Resolution Method | FSC 0.143 CUT-OFF |
| Other Details | |
| Refinement Type | |
| Symmetry Type | POINT |
| Point Symmetry | C4 |
| Map-Model Fitting and Refinement | |||||
|---|---|---|---|---|---|
| Id | 1 | ||||
| Refinement Space | REAL | ||||
| Refinement Protocol | AB INITIO MODEL | ||||
| Refinement Target | |||||
| Overall B Value | |||||
| Fitting Procedure | |||||
| Details | Model refined against the cryosparc b-factor sharpened map using default restraints | ||||
| Data Acquisition | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Detector Type | GATAN K2 SUMMIT (4k x 4k) | ||||||||
| Electron Dose (electrons/Å**2) | 48.7 | ||||||||
| Imaging Experiment | 1 |
|---|---|
| Date of Experiment | |
| Temperature (Kelvin) | |
| Microscope Model | FEI TITAN KRIOS |
| Minimum Defocus (nm) | 1300 |
| Maximum Defocus (nm) | 2500 |
| Minimum Tilt Angle (degrees) | |
| Maximum Tilt Angle (degrees) | |
| Nominal CS | |
| Imaging Mode | BRIGHT FIELD |
| Specimen Holder Model | FEI TITAN KRIOS AUTOGRID HOLDER |
| Nominal Magnification | 37313 |
| Calibrated Magnification | |
| Source | FIELD EMISSION GUN |
| Acceleration Voltage (kV) | 300 |
| Imaging Details | EBIC TITAN KRIOS M02 |
| EM Software | ||
|---|---|---|
| Task | Software Package | Version |
| PARTICLE SELECTION | Gautomatch | 0.53 |
| IMAGE ACQUISITION | EPU | |
| CTF CORRECTION | CTFFIND | 4.1.5 |
| MODEL FITTING | Coot | 0.8.9.1 |
| MODEL REFINEMENT | PHENIX | 1.13 |
| INITIAL EULER ASSIGNMENT | RELION | 2.1-beta-0 |
| FINAL EULER ASSIGNMENT | RELION | 2.1-beta-0 |
| CLASSIFICATION | RELION | 2.1-beta-0 |
| RECONSTRUCTION | cryoSPARC | 0.6.5 |
| Image Processing | ||||
|---|---|---|---|---|
| CTF Correction Type | CTF Correction Details | Number of Particles Selected | Particle Selection Details | |
| PHASE FLIPPING AND AMPLITUDE CORRECTION | 157681 | Picking with templates derived from manually picked particles | ||
