TABLE 1:
Cryo-EM data collection, refinement and validation statistics
| DUOX1–DUOXA1 (apo) (EMDB-21962, PDB 6WXR) | DUOX1–DUOXA1 (dimer of dimers) (EMDB-21963, PDB 6WXU) | DUOX1–DUOXA1 (with NADPH) (EMDB-21964, PDB 6WXV) | |
|---|---|---|---|
| Data collection and processing | |||
| Magnification | 81,000 | 81,000 | |
| Voltage (kV) | 300 | 300 | |
| Electron exposure (e–/Å2) | 65.4 | 80.5 | |
| Defocus range (μm) | 1.1–2.0 | 1.1–2.0 | |
| Pixel size (Å) | 1.08 | 1.08 | |
| Symmetry imposed | C1 | C2 | C1 |
| Initial particle images (no.) | ~2,200,000 | 1,486,046 | |
| Final particle images (no.) | 534,337 | 302,097 | 667,038 |
| Map resolution (Å) | 3.2 | 2.7 | 3.3 |
| FSC threshold | 0.143 | 0.143 | 0.143 |
| Map resolution range (Å) | 3.2–200 | 2.7–200 | 3.3–200 |
| Refinement | |||
| Initial model used (PDB code) | de novo | de novo | de novo |
| Model resolution (Å) | 3.2 | 2.7 | 3.3 |
| FSC threshold | 0.143 | 0.143 | 0.143 |
| Model resolution range (Å) | 3.2–200 | 2.7–200 | 3.3–200 |
| Map sharpening B factor (Å2) | −135 | −105 | −125 |
| Model composition | |||
| Non-hydrogen atoms | 10,410 | 18,419 | 10,610 |
| Protein residues | 1,303 | 2,212 | 1,319 |
| Ligands | 4 | 8 | 5 |
| B factors (Å2) | |||
| Protein | 36.06 | 67.61 | 41.73 |
| Ligand | - | - | 30.07 |
| R.m.s. deviations | |||
| Bond lengths (Å) | 0.003 | 0.006 | 0.004 |
| Bond angles (°) | 0.716 | 0.694 | 0.702 |
| Validation | |||
| MolProbity score | 1.90 | 1.71 | 1.82 |
| Clashscore | 8.68 | 3.62 | 8.23 |
| Poor rotamers (%) | 0.19 | 1.95 | 0 |
| Ramachandran plot | |||
| Favored (%) | 93.31 | 95.18 | 94.46 |
| Allowed (%) | 6.69 | 4.82 | 5.54 |
| Disallowed (%) | 0 | 0 | 0 |