Table 1.
Data collection and refinement statistics for GTB double and triple mutants –15/GTB/C80S/C196S and –15/GTB/C80S/C196S/C209S in their unliganded and acceptor-bound states
| Enzyme |
C80S/C196S |
C80S/C196S/C209S |
||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Substrates | Unliganded | H antigen | UDP | UDP + H | UDP-Gal + DA | Unliganded | H antigen | UDP | UDP + H | UDP-Gal + DA |
| Resolution (Å)a,b | 20–1.55 (1.61–1.55) | 20–1.81 (1.87–1.81) | 20–1.90 (1.97–1.90) | 20–1.56 (1.62–1.56) | 20–1.40 (1.45–1.40) | 20–1.79 (1.85–1.79) | 20–1.48 (1.53–1.48) | 20–1.49 (1.54–1.59) | 20–1.93 (2.00–1.93) | 20–1.60 (1.66–1.60) |
| No. of observations | 193,697 | 125,758 | 106,079 | 197,444 | 257,813 | 134,916 | 187,580 | 216,207 | 111,381 | 169,278 |
| Unique reflections | 43,878 | 27,621 | 24,723 | 43,699 | 58,949 | 28,441 | 51,095 | 49,738 | 22,845 | 39,424 |
| Completeness (%)a | 96.4 (98.1) | 95.6 (93.9) | 99.0 (99.7) | 97.3 (99.6) | 95.3 (76.9) | 96.2 (98.1) | 97.2 (99.4) | 96.7 (96.7) | 95.8 (98.5) | 94.6 (89.5) |
| Rmerge (%)a | 4.2 (28.8) | 7.4 (31.7) | 7.8 (31.4) | 4.1 (30.0) | 3.5 (32.5) | 5.1 (29.9) | 3.8 (24.9) | 3.5 (39.7) | 5.1 (29.8) | 4.6 (26.9) |
| I/σ(I) | 17.1 (3.8) | 12.0 (4.1) | 10.5 (4.2) | 17.3 (3.6) | 19.2 (3.2) | 14.0 (4.0) | 14.8 (4.1) | 17.2 (2.8) | 15.2 (4.1) | 15.3 (4.1) |
| Rwork (%) | 18.98 | 17.32 | 18.41 | 18.85 | 19.45 | 16.52 | 20.19 | 16.92 | 17.45 | 18.40 |
| Rfree (%) | 22.76 | 22.23 | 22.94 | 21.72 | 22.96 | 21.97 | 23.82 | 22.57 | 22.81 | 22.28 |
| RMS | ||||||||||
| Bond lengths (Å) | 0.028 | 0.025 | 0.025 | 0.030 | 0.030 | 0.023 | 0.027 | 0.028 | 0.023 | 0.030 |
| Bond angles (°) | 2.305 | 1.931 | 2.189 | 2.247 | 2.572 | 1.80 | 2.178 | 1.961 | 2.202 | 2.447 |
| Mean B (Å2) | 21.44 | 21.24 | 26.57 | 20.93 | 18.88 | 24.51 | 21.08 | 22.77 | 27.62 | 19.79 |
| Protein | 19.54 | 19.63 | 25.69 | 19.24 | 16.93 | 23.75 | 19.69 | 21.55 | 25.97 | 18.01 |
| Ligandsc | NA | 22.85 | 42.39 | 22.68 | 20.94 | NA | 22.49 | 39.98 | 33.92 | 20.89 |
| Waterc | 34.41 | 32.12 | 35.50 | 33.19 | 30.79 | 33.01 | 31.79 | 33.05 | 32.51 | 31.74 |
| No. of water | 320 | 301 | 210 | 306 | 344 | 202 | 269 | 227 | 143 | 317 |
| PDB ID | 3I0C | 3I0E | 3I0D | 3I0F | 3I0G | 3I0H | 3I0J | 3I0I | 3I0K | 3I0L |
a
Values in parentheses are for the highest-resolution shell.
b
All crystals belonged to space group C2221, with unit cell dimensions ranging from 52.1 to 52.9 Å, from 148.6 to 150.4 Å, and from 79.3 to 79.7 Å.
c
All ligand and water molecules are refined at 100% occupancy.