2PZN

The crystallographic structure of Aldose Reductase IDD393 complex confirms Leu300 as a specificity determinant

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	1999-05-27
Detector	SBC-2
Wavelength(s)	0.90042
Spacegroup name	P 1
Unit cell lengths	47.224, 47.154, 40.316
Unit cell angles	67.58, 76.47, 76.11

Refinement procedure

Resolution	10.000 - 1.000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	ALDOSE REDUCTASE COMPLEXED WITH IDDD552 (PDB ID: 1T41)
RMSD bond length	0.014
RMSD bond angle	0.033
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	SHELXL-97

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.040
High resolution limit [Å]	1.000	1.000
Rmerge	0.062	0.271
Number of reflections	158045
<I/σ(I)>	13.65	3.6
Completeness [%]	94.9	85.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	298	Protein was co-crystallized with NADP+ (Sigma) and inhibitor (ratios protein/coenzyme/inhibitor = 1/2/2). Previously equilibrated (ammonium citrate buffer, PEG 6000 (15%)) hanging drops were seeded with stock seed solutions diluted 100 times. Cryofreezing was carried out through quick transfers into a stabilization solution (25% PEG 6000), then into a cryo-protecting solution (40% PEG 6000) and finally into either liquid nitrogen or ethane, VAPOR DIFFUSION, HANGING DROP, temperature 298K