Table 1. Crystallization conditions and crystallographic parameters.
Values in parentheses are for the outermost resolution shell.
| Wild type | Tyr29Phe | Tyr29Ala | |
|---|---|---|---|
| Precipitant | 1.6M ammonium sulfate | 1.2M ammonium sulfate | 1.3M ammonium sulfate |
| Buffer | 0.05M Na2HP2O7 | 0.05M Na2HP2O7 | 0.05M Na2HP2O7 |
| pH | 6.0 | 6.0 | 6.2 |
| Cryoprotectant | 3% glycerol | 3% glycerol | 3% ethylene glycol |
| Protein concentration (mgml1) | 25 | 30 | 27 |
| Streak-seeded? | Yes | Yes | No |
| Crystal dimensions (mm) | 0.2 0.07 0.04 | 0.2 0.07 0.04 | 0.2 0.07 0.04 |
| Resolution range () | 30.161.75 (1.811.75) | 31.201.90 (1.971.90) | 30.951.72 (1.781.72) |
| Space group | C2221 | P21 | C2 |
| Unit-cell parameters | |||
| a () | 74.9 | 61.6 | 84.9 |
| b () | 98.9 | 41.1 | 46.3 |
| c () | 41.0 | 62.4 | 62.5 |
| () | 90 | 105.6 | 128.6 |
| No. of reflections | 15731 (1561) | 23653 (2082) | 22086 |
| Completeness (%) | 99.87 (100) | 98.39 (86.97) | 98.72 (95.95) |
| I/(I) | 22.53 (3.97) | 24.13 (7.84) | 15.12 (3.06) |
| R merge | 0.0564 (0.3741) | 0.0471 (0.1516) | 0.0730 (0.3768) |
| R | 0.1739 (0.2495) | 0.1571 (0.1749) | 0.1712 (0.3143) |
| R free | 0.2092 (0.2561) | 0.1916 (0.2245) | 0.2040 (0.3347) |
| Protein atoms per monomer | 1147 | 1122 | 1161 |
| Waters per monomer | 72 | 80 | 86 |
| Cryoprotectant molecules per monomer | 6 | 5.5 | 14 |
| Alternate conformers† | 14 | 10 | 13 |
| R.m.s.d., bond lengths () | 0.004 | 0.013 | 0.018 |
| R.m.s.d., bond angles () | 1.06 | 1.56 | 1.51 |
| Ramachandran favored‡ (%) | 98 | 97 | 99 |
| Ramachandran outliers‡ (%) | 0.72 | 0 | 0 |
| Solvent content (%) | 48.5 | 48.7 | 55.6 |
| B, main chain (2) | 33.9 | 24.8 | 23.0 |
| B, side chains (2) | 41.8 | 32.3 | 28.8 |
| B, solvent§ (2) | 41.2 | 30.3 | 39.9 |
| No. of residues in dimer interface | n.a. | 22 | n.a. |
| PDB code | 3tm3 | 3tld | 3tm9 |
†
Number of protein residues per monomer that were built in two or more conformations.
‡
Number of residues per monomer in the favored or allowed region of the Ramachandran plot.
§
Including glycerol or ethylene glycol atoms.