1W5P
Stepwise introduction of zinc binding site into porphobilinogen synthase of Pseudomonas aeruginosa (mutations A129C, D131C, D139C, P132E)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | MPG/DESY, HAMBURG BEAMLINE BW6 |
Synchrotron site | MPG/DESY, HAMBURG |
Beamline | BW6 |
Temperature [K] | 100 |
Detector technology | IMAGE PLATE |
Collection date | 2003-11-04 |
Detector | MARRESEARCH |
Spacegroup name | P 4 21 2 |
Unit cell lengths | 126.970, 126.970, 86.432 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 91.290 - 1.550 |
R-factor | 0.159 |
Rwork | 0.157 |
R-free | 0.18500 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1b4k |
RMSD bond length | 0.015 |
RMSD bond angle | 1.573 |
Data reduction software | DENZO |
Data scaling software | SCALEPACK |
Phasing software | CNS |
Refinement software | REFMAC (5.1.24) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 20.000 | 1.610 |
High resolution limit [Å] | 1.550 | 1.550 |
Rmerge | 0.081 | 0.500 |
Number of reflections | 101482 | |
<I/σ(I)> | 16 | 2 |
Completeness [%] | 99.0 | 99 |
Redundancy | 6.7 | 6.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | SITTING DROP. DROPS WERE MIXED OF 3 MICROLITER OF PROTEIN SOLUTION (12 MG/ML PROTEIN, 50 MM NA-HEPES PH 7.5, 10MM MGCL2, 1MM ZNCL2, 10 MM DTT) PLUS 3 MICROLITER OF RESERVOIR SOLUTION (1M LITHIUM SULFATE, 100MM TRIS-HCL PH 8.5, 10MM NICKEL CHLORIDE), 100 MICROLITER OF RESERVOIR SOLUTION, 96 WELL PLATES (NUNC) IN AN ANAEROBIC BOX |