Table 1 Cryo-EM data collection, refinement and validation statistics
Native WT Polymorph 1 (EMDB-61782) (PDB 9JST) | Native WT Polymorph 2 (EMDB-61783) (PDB 9JSU) | Native G175S (EMDB-61784) (PDB 9JSV) | In vitro WT (EMDB-61785) (PDB 9JSW) | In vitro G175S (EMDB-61786) (PDB 9JSX) | |
|---|---|---|---|---|---|
Data collection and processing | |||||
Magnification | 60,000 | 60,000 | 60,000 | 60,000 | 60,000 |
Voltage (kV) | 300 | 300 | 300 | 300 | 300 |
Electron exposure (e–/Å2) | 50 | 50 | 50 | 50 | 50 |
Defocus range (μm) | 0.8–2.0 | 0.8–2.0 | 0.8–2.0 | 0.8–2.0 | 0.8–2.0 |
Pixel size (Å) | 0.8784 | 0.8784 | 0.8784 | 0.8784 | 0.8784 |
Symmetry imposed | Helical | Helical | Helical | Helical | Helical |
twist: 177.7° | twist: −4.1° | twist: 178.4° | twist: 177.8° | twist: 178.4° | |
rise: 2.34 Å | rise: 4.67 Å | rise: 2.35 Å | rise: 2.31 Å | rise: 2.38 Å | |
Initial particle images (no.) | 30,430,102 | 30,430,102 | 6,395,243 | 3,826,618 | 3,606,507 |
Final particle images (no.) | 286,491 | 352,878 | 576,997 | 147,735 | 781,124 |
Map resolution (Å) | 1.79 | 1.79 | 1.79 | 1.94 | 1.79 |
FSC threshold | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 |
Map resolution range (Å) | 1.79–1.95 | 1.79–1.95 | 1.79–1.95 | 1.94–2.1 | 1.79–1.95 |
Refinement | |||||
Initial model used (PDB code) | ModelAngelo | ModelAngelo | ModelAngelo | ModelAngelo | ModelAngelo |
Model resolution (Å) | 1.7 | 1.7 | 1.7 | 2.0 | 1.8 |
FSC threshold | 0.143 | 0.143 | 0.143 | 0.143 | 0.143 |
Model resolution range (Å) | 1.7–2.0 | 1.7–2.0 | 1.7–2.0 | 2.0–2.1 | 1.7–2.0 |
Map sharpening B factor (Å2) | −272 | −255 | −268 | −268 | −272 |
Model composition | |||||
Non-hydrogen atoms | 538 | 505 | 531 | 538 | 531 |
Protein residues | 35 | 33 | 34 | 35 | 34 |
Ligands | 0 | 0 | 0 | 0 | 0 |
B factors (Å2) | |||||
Protein | 18.17/55.61/30.58 | 63.33/100.00/98.61 | 80.76/100.00/98.81 | 18.17/55.61/30.58 | 80.76/100.0/98.81 |
R.m.s. deviations | |||||
Bond lengths (Å) | 0.090 | 0.089 | 0.090 | 0.089 | 0.088 |
Bond angles (°) | 1.882 | 1.876 | 1.884 | 1.880 | 1.985 |
Validation | |||||
MolProbity score | 0.91 | 0.70 | 0.69 | 0.91 | 0.91 |
Clashscore | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 |
Poor rotamers (%) | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 |
Ramachandran plot | |||||
Favored (%) | 93.94 | 96.77 | 96.88 | 93.94 | 93.75 |
Allowed (%) | 6.06 | 3.23 | 3.12 | 6.06 | 6.25 |
Disallowed (%) | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 |
