3T42
Human aldose reductase in complex with a nitrile-containing IDD inhibitor
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SSRL BEAMLINE BL9-2 |
Synchrotron site | SSRL |
Beamline | BL9-2 |
Temperature [K] | 93 |
Detector technology | CCD |
Collection date | 2010-04-20 |
Detector | MARMOSAIC 325 mm CCD |
Spacegroup name | P 1 |
Unit cell lengths | 39.858, 46.963, 46.907 |
Unit cell angles | 76.35, 77.21, 67.79 |
Refinement procedure
Resolution | 22.290 - 1.280 |
R-factor | 0.152 |
Rwork | 0.151 |
R-free | 0.17800 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 2pzn |
RMSD bond length | 0.008 |
RMSD bond angle | 1.341 |
Data reduction software | HKL-2000 |
Data scaling software | SCALEPACK |
Phasing software | PHASER |
Refinement software | PHENIX ((phenix.refine: 1.6_289)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 50.000 | 1.330 |
High resolution limit [Å] | 1.280 | 1.280 |
Rmerge | 0.042 | 0.301 |
Number of reflections | 73538 | |
<I/σ(I)> | 18.4 | |
Completeness [%] | 94.3 | 87.7 |
Redundancy | 3.9 | 3.4 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | 4.2 | 298 | The hanging drop method was used to obtain crystals (EasyXtal Tool 24 culture plates, Qiagen). WT hALR2 was co-crystallized with the oxidized form of coenzyme NADP+ (Sigma) and inhibitor 2 at room temperature (ratios of protein / inhibitor / coenzyme = 1/2/2). Hanging drops were made by the mixing mother liquor (0.06 M citric acid, 0.04 M Bis-tris propane, 12% (w/v) PEG 3350, pH 4.2) with holoenzyme solution (10 mg/mL, 20 mM hepes, pH 7.0). Crystals were observed after three days of equilibration, EVAPORATION, temperature 298K |