Table 1.
CryoEM data collection, refinement and validation statistics
| Dantrolene/ATP/4CmC/Caffeine/Mg2+ | Dantrolene/ADP/4CmC/Caffeine/Mg2+ | Fatigue | Physiological | ATP/4CmC/Caffeine | ADP/4CmC/Caffeine/Mg2+ | Dantrolene/Mg2+ | ATP/Mg2+/Ca2+ | |
|---|---|---|---|---|---|---|---|---|
| Data Collection | ||||||||
| Magnification | 130,000 x | 130,000 x | 130,000 x | 130,000 x | 130,000 x | 130,000 x | 130,000 x | 130,000 x |
| Pixel size (Å/pix) | 1.06 | 0.83 | 0.83 | 0.83 | 1.06 | 0.83 | 0.83 | 0.83 |
| Voltage (kV) | 300 | 300 | 300 | 300 | 300 | 300 | 300 | 300 |
| Total Electron exposure (e−/Å2) | 57 | 58.2 | 58 | 58 | 71.2 | 58 | 58 | 58 |
| Exposure rate (e−/pixel/seconds) | 8 | 16 | 16 | 16 | 8 | 16 | 16 | 16 |
| Number of frames per movie | 40 | 50 | 50 | 50 | 50 | 50 | 50 | 50 |
| Defocus range (μm) | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 | 0.5–1.5 |
| Initial micrographs | 5,959 | 3,928 | 5,405 | 3,556 | 8,271 | 9,130 | 3,954 | 4,975 |
| Final micrographs | 4,897 | 3,084 | 5,228 | 3,105 | 7,302 | 8,493 | 1,584 | 4,440 |
| Data Processing | ||||||||
| Initial particle images | 148,129 | 145,871 | 381,615 | 331,769 | 470,981 | 808,919 | 59,519 | 145,024 |
| Final particle images | 71,984 | 66,232 | 224,598 | 177,397 | 259,362 | 294,178 | 28,581 | 82,245 |
| Box size (unbinned) (pixels) | 512 | 600 | 600 | 450 (600) | 512 | 600 | 450 (600) | 600 |
| Symmetry imposed | C4 | C4 | C4 | C4 | C4 | C4 | C4 | C4 |
| Map resolution (FSC at 0.143) | ||||||||
| Masked (Å) | 3.1 | 2.6 | 2.2 | 2.7 | 2.8 | 2.4 | 3.0 | 2.6 |
| Unmasked (Å) | 3.9 | 3.4 | 3.0 | 3.3 | 3.3 | 3.0 | 4.0 | 3.2 |
| Map sharpening B factor (Å2) | 77.6 | 59.8 | 55.2 | 78.4 | 88.5 | 66.4 | 53.3 | 64.5 |
| Modeling | ||||||||
| Model composition | ||||||||
| Non-H atoms | 140,460 | 140,583 | 143,344 | 143,036 | 143,361 | 143,832 | 142,668 | 143,440 |
| Protein residues | 21,288 | 21,300 | 21,288 | 21,288 | 21,288 | 21,288 | 21,296 | 21,288 |
| R.m.s. deviations | ||||||||
| Bond lengths (Å) | 0.26 | 0.26 | 0.30 | 0.25 | 0.27 | 0.26 | 0.27 | 0.27 |
| Bond angles (°) | 0.48 | 0.48 | 0.51 | 0.49 | 0.50 | 0.48 | 0.53 | 0.49 |
| Validation | ||||||||
| Clashscore | 5 | 5 | 5 | 6 | 5 | 6 | 11 | 7 |
| Rotamer outliers (%) | 2 | 4 | 3 | 4 | 4 | 3 | 6 | 4 |
| Ramachandran plot | ||||||||
| Favored (%) | 99 | 99 | 98 | 98 | 98 | 98 | 98 | 98 |
| Allowed (%) | 1 | 1 | 2 | 2 | 2 | 2 | 2 | 2 |
| Outliers (%) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| PDB code | xxxx | xxxx | xxxx | xxxx | xxxx | xxxx | xxxx | xxxx |
| EMDB code | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx | EMD-xxxxx |
| EMPIAR code | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx | EMPIAR-xxxxx |