3SEF

2.4 Angstrom resolution crystal structure of shikimate 5-dehydrogenase (aroE) from Vibrio cholerae O1 biovar eltor str. N16961 in complex with shikimate and NADPH

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	2010-11-08
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97872
Spacegroup name	P 1 21 1
Unit cell lengths	76.030, 86.079, 81.018
Unit cell angles	90.00, 92.85, 90.00

Refinement procedure

Resolution	29.480 - 2.400
R-factor	0.23666
Rwork	0.235
R-free	0.27560
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3pgj
RMSD bond length	0.005
RMSD bond angle	1.267
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.440
High resolution limit [Å]	2.400	2.400
Rmerge	0.055	0.570
Number of reflections	41040
<I/σ(I)>	18.92	2.31
Completeness [%]	99.9	100
Redundancy	3.8	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	295	Protein: 6.8 mg/mL in 10 mM Tris/HCl pH 8.3, 0.25 M NaCl, 5 mM BME. Soak: 5 mM NADPH, 5 mM dehydroshikimate. Crystallization: PEGsII (B11: 0.2 M Na acetate, 0.1 M HEPES pH 7.5, 20 % (w/v) PEG3000, VAPOR DIFFUSION, SITTING DROP, temperature 295K