Extended Data Table 1.
Cryo-EM data collection, refinement and validation statistics.
| Full-length NKCC1 | N-terminally truncated NKCC1 |
|||||
|---|---|---|---|---|---|---|
| Data collection and processing | ||||||
| Magnification | 130,000 | 130,000 | ||||
| Voltage (kV) | 300 | 300 | ||||
| Electron exposure (e−/Å2) | 53 | 53 | ||||
| Defocus range (μm) | 1.0–3.0 | 1.0–2.5 | ||||
| Pixel size (Å) | 1.06 | 1.04 | ||||
| Exposure time (seconds) | 8 | 10 | ||||
| Number of movies | 4,586 | 5,845 | ||||
| Initial particle images (no.) | 1,005,359 | 914,197 | ||||
| TM domain |
CTD | Composite TM+CTD |
Full Molecule |
TM domain | CTD | |
| PDB 6NPH EMDB 0470 | PDB 6NPJ EMDB 0471 | PDB 6NPL EMDB 0473 | EMDB 0475 | PDB 6NPK EMDB 0472 | EMDB 0474 | |
| Symmetry imposed | C2 | C1 | C1 | C2 | Cl | |
| Final particle images (no.) | 63,659 | 60,997 | 60,997 | 227,231 | 233,031 | |
| Map resolution (Å) FSC threshold |
2.9 0.143 |
3.8 0.143 |
4.2 0.143 |
3.6 0.143 |
4.2 0.143 |
|
| Map resolution range (Å) | 3.9–2.9 | 6.6–3.7 | 8.8–4.0 | 6.2–3.4 | 7.0–4.0 | |
| Refinement | ||||||
| Initial model used (PDB code) | Individual TM domain and CTD | |||||
| Model resolution (Å) FSC threshold |
3.1 0.5 |
3.8 0.5 |
3.8 0.5 |
|||
| Model resolution range (Å) | 233–2.9 | 233–3.8 | 233–2.9 | |||
| Map sharpening B factor (Å2) | −107 | −162 | −191 | −209 | −250 | |
| Model composition | ||||||
| Non-hydrogen atoms | 7504 | 6186 | 13774 | 7134 | ||
| Protein residues | 944 | 768 | 1722 | 944 | ||
| Ligands | 13 | -- | 13 | -- | ||
| B factors (Å2) | ||||||
| Protein | 64.27 | 39.82 | 31.72 | 57.11 | ||
| Ligand | 65.84 | -- | 22.94 | -- | ||
| R.m.s. deviations | ||||||
| Bond lengths (Å) | 0.008 | 0.008 | 0.006 | 0.004 | ||
| Bond angles (°) | 1.090 | 1.236 | 0.977 | 0.820 | ||
| Validation | ||||||
| MolProbity score | 2.26 | 2.47 | 2.62 | 2.21 | ||
| Clashscore | 5.09 | 6.43 | 9.03 | 3.89 | ||
| Poor rotamers (%) | 4.27 | 6.25 | 7.74 | 6.13 | ||
| Ramachandran plot | ||||||
| Favored (%) | 92.13 | 91.18 | 91.48 | 93.40 | ||
| Allowed (%) | 7.87 | 8.82 | 8.46 | 6.60 | ||
| Disallowed (%) | 0 | 0 | 0.06 | 0 | ||