1R6W
Crystal structure of the K133R mutant of o-Succinylbenzoate synthase (OSBS) from Escherichia coli. Complex with SHCHC
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 14-BM-D |
Synchrotron site | APS |
Beamline | 14-BM-D |
Temperature [K] | 150 |
Wavelength(s) | 0.9790 |
Spacegroup name | P 21 21 2 |
Unit cell lengths | 72.200, 82.900, 56.200 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 30.000 - 1.620 |
R-factor | 0.1691 |
Rwork | 0.167 |
R-free | 0.20000 * |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1fhv |
RMSD bond length | 0.016 |
RMSD bond angle | 1.620 * |
Data reduction software | HKL-2000 |
Data scaling software | SCALEPACK |
Phasing software | MOLREP |
Refinement software | REFMAC (5.1.24) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 31.300 * | 1.680 |
High resolution limit [Å] | 1.620 | 1.620 |
Rmerge | 0.046 | 0.293 |
Total number of observations | 307914 * | |
Number of reflections | 43694 | |
<I/σ(I)> | 41 | 17.1 |
Completeness [%] | 99.8 * | 99.9 * |
Redundancy | 7.05 * |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | Batch method * | 5.5 | 20 * | The K133R mutant of OSBS was concentrated to 15 mg/ml, dialyzed against 5 mM Tris pH 8.3 containing 2 mM MgCl2, drop frozen as small pellets in liquid nitrogen and stored at -80 C. Crystals were grown at 20 C by small-scale batch experiments by combining 15 ml of protein solution and 15 ml of a solution containing 12-13% MePEG 5000, 100 mM sodium acetate, 60 mM MgCl2, at pH 5.5. SHCHC was included in the crystallization at a final concentration of approximately 2.5 mM., microbatch, temperature 293K |
Crystallization Reagents in Literatures
ID | crystal ID | solution | reagent name | concentration (unit) | details |
1 | 1 | 1 | protein | 15 (mg/ml) | |
2 | 1 | 1 | Tris-HCl | 5 (mM) | pH8.3 |
3 | 1 | 1 | 2 (mM) | ||
4 | 1 | 1 | MePEG5000 | 12-13 (%) | |
5 | 1 | 1 | sodium acetate | 100 (mM) | |
6 | 1 | 1 | 60 (mM) | pH5.5 |