Table 2.
Wild type | Wild type + pyruvate | E192N | E192N + pyruvate | E192N + pyuvate + THB | |
---|---|---|---|---|---|
Diamond beamline | I02 | I03 | I04 | I03 | I03 |
Space group | P21 | P21 | P21 | P21 | P21 |
a, b, c (Å), | 54.8, 142.2, 84.2 | 54.7, 142.5, 83.6 | 54.6, 142.8, 84.5 | 56.9, 143.0, 83.9 | 57.0, 143.7, 83.3 |
β (°) | 109.0 | 109.2 | 109.0 | 109.8 | 109.9 |
Resolution (Å) | 79.56–2.20 (2.26–2.20) | 79.1–1.65 (1.74–1.65) | 47.60–1.80 (1.90–1.80) | 79.06–1.80 (1.95–1.80) | 79.31–2.05 (2.16–2.05) |
Rmerge | 0.102 (0.365) | 0.060 (0.317) | 0.086 (0.388) | 0.070 (0.430) | 0.09 (0.438) |
Rpim (all I+ and I−) | 0.063 (0.219) | 0.038 (0.215) | 0.054 (0.033) | 0.043 (0.269) | 0.056 (0.273) |
〈I〉/σ〈I〉 | 8.5 (3.4) | 13.5 (3.2) | 8.3 (2.8) | 12.0 (2.5) | 8.6 (2.6) |
Completeness (%) | 99.9 (100) | 91.2 (60.8) | 98. 4 (97.2) | 100 (99.9) | 98.6 (96.3) |
Redundancy | 3.7 (3.7) | 3.4 (2.9) | 3.6 (3.5) | 3.6 (3.5) | 3.5 (3.4) |
Wilson B-factor (Å2) | 34.8 | 31.7 | 26.8 | 21.8 | 24.5 |
Number of reflections | 197,621 (29,177) | 449,770 (37,158) | 398,910 (55,837) | 391,185 (54,278) | 277,866 (37,606) |
Number of unique reflections | 54,006 (7847) | 131,694 (12,746) | 110,985 (15,919) | 107,356 (15,671) | 78,597 (11,141) |
Resolution (Å) | 79.56–2.20 (2.26–2.20) | 79.1–1.65 (1.69–1.65) | 47.60–1.80 (1.90–1.80) | 79.06–1.80 (1.95–1.80) | 79.31–2.05 (2.16–2.05) |
R-factor | 0.211 (0.259) | 0.209 (0.261) | 0.197 (0.261) | 0.188 (0.301) | 0.190 (0.281) |
Rfree | 0.271 (0.356) | 0.247 (0.344) | 0.253 (0.391) | 0.225 (0.344) | 0.234 (0.337) |
Twinning fraction | 0.338 | 0.334 | 0.463 | None | 0.06 |
Twinning operator | − h, − k, h + l | − h, − k, h + l | − h, − k, h + l | None | − h, − k, h + l |
Number of atoms | |||||
Protein | 9166 | 9166 | 9178 | 9292 | 9246 |
Water | 169 | 758 | 316 | 654 | 540 |
Ligands | — | 31 (PEG 400, Na+) | 4 (Cl) | 24 (PEG 400, pyruvatea) | 60 (THB) |
Average B-factors (Ǻ2) | |||||
Protein | 32.9 | 22.8 | 28.0 | 22.8 | 28.4 |
Waters | 27.9 | 26.7 | 26.7 | 27.2 | 31.9 |
Ligands | — | 32.4 | 26.3 | 45.8 | 50.1 |
RMSDs | |||||
Bond lengths (Å) | 0.013 | 0.012 | 0.011 | 0.009 | 0.012 |
Bond angles (°) | 1.46 | 1.80 | 1.33 | 1.14 | 1.28 |
Ramachandran plot, most favored region (%) | 94.2b | 98.3b | 97.6b | 98.0b | 98.3b |
PDB code | 2WO5 | 2WNN | 2WNQ | 2WNZ | 2WPB |
Two additional molecules of noncovalently bound pyruvate are observed in subunits C and D.
In all structures, 100% of residues lie within favored regions of the Ramachandran plot.