4XAP

Crystal structure of aldo-keto reductase from Sinorhizobium meliloti 1021

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2014-11-06
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.978
Spacegroup name	P 21 21 21
Unit cell lengths	44.533, 78.068, 79.073
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 2.210
R-factor	0.1634
Rwork	0.160
R-free	0.22460
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4pmj
RMSD bond length	0.015
RMSD bond angle	1.720
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0049)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.240
High resolution limit [Å]	2.200	5.970	2.200
Rmerge	0.072	0.037	0.692
Rmeas	0.080	0.042	0.768
Rpim	0.035	0.019	0.327
Total number of observations	65764
Number of reflections	14216
<I/σ(I)>	17.6		2.1
Completeness [%]	98.7	95.2	98.7
Redundancy	4.6	4.3	4.5
CC(1/2)		0.998	0.805

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	289	0.2 ul of 15 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide and 0.5 mM TCEP were mixed with 0.2 ul of the PEG/Ion HT condition #70 (0.2M Ammonium Citrate Tribasic anhydrous, 20% w/v PEG 3350 pH=7) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 1/50 v/v of 1 mg/ml TEV solution at 289 K for 1 hour