Table 1.
Crystallographic data for thrombin mutants N143P and Y225P.
| N143P - E form | N143P - E* form | Y225P - E form | Y225P - E* form | |
|---|---|---|---|---|
| Buffer/salt | 0.2 M NH4I | 0.1 M imidazole | 0.2 M K formate | 0.1 M Tris, pH 8.0 |
| PEG | 3350 (20%) | 8000 (7%) | 3350 (20%) | 8000 (8%) |
| PDB ID | 3QGN | 3JZ2 | 3S7K | 3S7H |
|
| ||||
| Data collection: | Raxis IV++ | Mar345 | Raxis IV++ | Raxis IV++ |
| Wavelength (Å) | 1.54 | 1.54 | 1.54 | 1.54 |
| Space group | C2 | P43 | P21212 | P43 |
| Unit cell dimensions (Å) | a=122.2 | a=57.9 | a=61.9 | a=57.6 |
| b=48.0 | b=57.9 | b=86.8 | b=57.6 | |
| c=52.0 | c=119.8 | c=101.0 | c=119.9 | |
| β=94.3° | ||||
| Molecules/asymmetric unit | 1 | 1 | 2 | 1 |
| Resolution range (Å) | 40-2.1 | 40-2.4 | 40-1.9 | 40-1.9 |
| Observations | 75156 | 83891 | 286602 | 185752 |
| Unique observations | 17583 | 14942 | 43419 | 30093 |
| Completeness (%) | 98.5 (97.3) | 96.7 (77.8) | 99.3 (98.2) | 97.8 (86.3) |
| Rsym (%) | 10.5 (16.0) | 7.6 (40.8) | 5.4 (30.5) | 7.3 (41.8) |
| I/σ(I) | 13.3 (6.6) | 19.0 (2.2) | 27.6 (4.0) | 20.7 (2.4) |
|
| ||||
| Refinement: | ||||
| Resolution (Å) | 40-2.1 | 40-2.4 | 40-1.9 | 40-1.9 |
| Rcryst, Rfree | 0.189, 0.239 | 0.189, 0.246 | 0.175, 0.216 | 0.175, 0.204 |
| Reflections (working/test) | 15588/892 | 14167/747 | 38897/2179 | 27031/1518 |
| Protein atoms | 2253 | 2242 | 4589 | 2273 |
| Solvent molecules | 150 | 116 | 382 | 191 |
| Rmsd bond lengthsa (Å) | 0.010 | 0.012 | 0.010 | 0.0080 |
| Rmsd anglesa (°) | 1.3 | 1.4 | 1.3 | 1.1 |
| Rmsd ΔB (Å2) (mm/ms/ss)b | 2.99/2.28/3.26 | 2.02/1.36/2.45 | 2.18/1.83/2.70 | 2.86/2.33/2.56 |
| <B> protein (Å2) | 41.8 | 34.1 | 35.4 | 39.0 |
| <B> solvent (Å 2) | 52.1 | 20.3 | 45.2 | 52.6 |
|
| ||||
| Ramachandran plot: | ||||
| Most favored (%) | 99.2 | 100 | 99.6 | 100 |
| Generously allowed (%) | 0.4 | 0 | 0.2 | 0 |
| Disallowed (%) | 0.4 | 0 | 0.2 | 0 |
a
Root-mean-squared deviation (Rmsd) from ideal bond lengths and angles and Rmsd in B-factors of bonded atoms.
b
mm, main chain-main chain; ms, main chain-side chain; ss, side chain-side chain. The structure of N143P in the E* form is from ref (29).