7PUO
Structure of a fused 4-OT variant engineered for asymmetric Michael addition reactions
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE MASSIF-1 |
| Synchrotron site | ESRF |
| Beamline | MASSIF-1 |
| Temperature [K] | 100 |
| Detector technology | PIXEL |
| Collection date | 2021-06-19 |
| Detector | DECTRIS PILATUS3 2M |
| Wavelength(s) | 0.965459 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 51.860, 100.915, 153.117 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 46.130 - 2.350 |
| R-factor | 0.2017 |
| Rwork | 0.200 |
| R-free | 0.23590 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 4x19 |
| RMSD bond length | 0.006 |
| RMSD bond angle | 0.849 |
| Data reduction software | XDS (20210205) |
| Data scaling software | Aimless (0.7.7) |
| Phasing software | PHASER (2.7.0) |
| Refinement software | PHENIX (1.20rc1-4395) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 47.920 | 47.920 | 2.430 |
| High resolution limit [Å] | 2.350 | 9.100 | 2.350 |
| Rmerge | 0.071 | 0.040 | 0.963 |
| Rmeas | 0.079 | 0.044 | 1.072 |
| Rpim | 0.034 | 0.019 | 0.459 |
| Total number of observations | 182172 | 3232 | 17108 |
| Number of reflections | 34025 | 663 | 3229 |
| <I/σ(I)> | 13.3 | 37.9 | 1.7 |
| Completeness [%] | 99.3 | 97.5 | 98.8 |
| Redundancy | 5.4 | 4.9 | 5.3 |
| CC(1/2) | 0.998 | 0.998 | 0.622 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 7 | 293 | Purified enzyme (16 mg/ml in 5 mM potassium phosphate buffer, pH 7.3) was crystallised in drops prepared by 1:1 mixing with a crystallization reservoir solution containing 20% PEG 2000, 10 mM urea in 0.1 M Bis-Tris, pH 7.0. |
