4KE6
Crystal structure D196N mutant of Monoglyceride lipase from Bacillus sp. H257 in complex with 1-rac-lauroyl glycerol
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | ROTATING ANODE |
Source details | BRUKER AXS MICROSTAR |
Temperature [K] | 100 |
Detector technology | IMAGE PLATE |
Collection date | 2013-03-22 |
Detector | MAR scanner 345 mm plate |
Wavelength(s) | 1.54 |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 39.185, 182.878, 248.264 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 61.357 - 2.800 |
R-factor | 0.2021 |
Rwork | 0.200 |
R-free | 0.24550 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.008 |
RMSD bond angle | 1.360 |
Data reduction software | MOSFLM |
Data scaling software | SCALA (3.3.21) |
Phasing software | PHASER (2.5.2) |
Refinement software | PHENIX (1.8_1069) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 73.621 | 73.621 | 2.950 |
High resolution limit [Å] | 2.800 | 8.850 | 2.800 |
Rmerge | 0.034 | 0.401 | |
Number of reflections | 42482 | ||
<I/σ(I)> | 9.4 | 15.8 | 1.9 |
Completeness [%] | 94.4 | 89.2 | 87.6 |
Redundancy | 3.9 | 4 | 3.1 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 6.9 | 293 | ~0.9 mM of bMGL(D196N) was mixed with 180 mM 1-(rac)-lauroyl glycerol dissolved in 99% EtOH to achieve a final protein-ligand ratio of 1:5. The protein-ligand mixture was incubated at 4 C for 1 hour. Initial crystals were obtained using the Morpheus screen in condition A4. These crystals were used for preparing a seed stock. The optimised crystals were obtained in a drop containing 0.9 mM bMGL(D196N); 56 % v/v MPD, 0.1 M HEPES pH 6.9 and 1:1000 dilution of seed stock in a ratio of 2:2:1 respectively. 1-LG powder was added to these crystallization drops and the crystals were soaked for 8h., VAPOR DIFFUSION, HANGING DROP, temperature 293K |