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