Table 1.
CryoEM Data Collection and Refinement Statistics
| IR/insulin A- V3E |
IR/insulin A- L13R (symmetric) |
IR/insulin A- L13R (asymmetric) |
IR/insulin B- L17R (symmetric) |
IR/insulin B- L17R (asymmetric) |
IR/insulin A- V3E/insulin A- L13R |
IR/unsaturated insulin WT 2 insulins bound (symmetric) |
IR/unsaturated insulin WT 1 insulin bound (asymmetric) |
IR/unsaturated insulin WT 2 insulins bound (asymmetric, conformation 1) |
IR/unsaturated insulin WT 2 insulins bound (asymmetric, conformation 2) |
|
|---|---|---|---|---|---|---|---|---|---|---|
| EMD-25188 PDB: 7SL1 |
EMD-25190 PDB:7SL3 |
EMD-25189 PDB:7SL2 |
EMD-25192 PDB:7SL6 |
EMD-25191 PDB:7SL4 |
EMD-25193 PDB:7SL7 |
EMD-25428 PDB:7STH |
EMD-25429 PDB:7STI |
EMD-25430 PDB:7STJ |
EMD-25431 PDB:7STK |
|
| Data collection and processing | ||||||||||
| Magnification | 60,241 | 46,296 | 46,296 | 46,296 | 46,296 | 60,241 | 46,296 | 46,296 | 46,296 | 46,296 |
| Voltage (kV) | 300 | 300 | 300 | 300 | 300 | 300 | 300 | 300 | 300 | 300 |
| Electron exposure (e−/Å2) | 60 | 60 | 60 | 60 | 60 | 60 | 60 | 60 | 60 | 60 |
| Defocus range (μm) | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 | 1.6 – 2.6 |
| Pixel size (Å) | 0.83 | 1.08 | 1.08 | 1.08 | 1.08 | 0.83 | 1.08 | 1.08 | 1.08 | 1.08 |
| Symmetry imposed | C2 | C2 | C1 | C2 | C1 | C2 | C2 | C1 | C1 | C1 |
| Initial particle images (no.) | 589,579 | 1,118,695 | 1,118,695 | 1,704,973 | 1,704,973 | 1,023,259 | 2,030,596 | 2,030,596 | 2,030,596 | 2,030,596 |
| Final particle images (no.) | 39,125 | 104,877 | 261,699 | 82,586 | 115,018 | 106,909 | 66,602 | 6,130 | 12,430 | 46,052 |
| Map resolution (Å) | 3.4 | 3.4 | 3.6 | 3.7 | 5 | 3.1 | 3.5 | 4.9 | 4.4 | 4 |
| FSC threshold | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 |
| Refinement | ||||||||||
| Initial model used (PDB code) | 4ZXB | 6PXV | 6PXV, 4ZXB | 6PXV | 6PXV, 4ZXB | 6PXV | 6PXV | 6PXV | 6PXV | 6PXV |
| Model composition | ||||||||||
| Nonhydrogen atoms | 13,892 | 14,008 | 14,517 | 14,008 | 13,558 | 14,778 | 13,996 | 13,531 | 13,515 | 13,911 |
| Protein residues | 1,722 | 1,734 | 1,799 | 1,734 | 1,676 | 1,830 | 1,734 | 1,678 | 1,672 | 1,726 |
| Ligands | ||||||||||
| R.m.s. deviations | ||||||||||
| Bond lengths (Å) | 0.002 | 0.005 | 0.004 | 0.002 | 0.003 | 0.004 | 0.003 | 0.004 | 0.005 | 0.004 |
| Bond angles (°) | 0.624 | 0.7 | 0.589 | 0.577 | 0.607 | 0.697 | 0.544 | 0.659 | 0.696 | 0.572 |
| Validation | ||||||||||
| MolProbity score | 1.97 | 1.89 | 1.99 | 1.73 | 2.09 | 2.03 | 1.94 | 2.38 | 2.58 | 2.15 |
| Clashscore | 9.72 | 8.63 | 11.53 | 6.49 | 15.24 | 10.88 | 9.00 | 25.44 | 35.69 | 15.08 |
| Poor rotamers (%) | 0.06 | 0 | 0 | 0 | 0.07 | 0 | 0 | 0 | 0 | 0.06 |
| Ramachandran plot | ||||||||||
| Favored (%) | 92.79 | 93.42 | 93.75 | 94.59 | 93.98 | 92.35 | 92.64 | 92.12 | 90.10 | 92.64 |
| Allowed (%) | 7.21 | 6.46 | 6.25 | 5.18 | 5.9 | 7.49 | 7.36 | 7.75 | 9.84 | 7.07 |
| Disallowed (%) | 0 | 0.12 | 0 | 0.24 | 0.12 | 0.17 | 0 | 0.12 | 0.06 | 0.3 |