3GTE

Crystal Structure of Dicamba Monooxygenase with Non-heme Iron

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2005-06-29
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.000
Spacegroup name	P 32
Unit cell lengths	81.110, 81.110, 158.810
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	20.000 - 1.950
Rwork	0.235
R-free	0.26170
Structure solution method	MR WAS PERFORMED USING A PRELIMINARY DMO STRUCTURE. THIS EARLY DMO STRUCTURE WAS OBTAINED LARGELY USING A HIGH-REDUNDANCY, 1.5418 A WAVELENGTH DATA SET FOR FE SINGLE ANOMALOUS DISPERSION (SAD) PHASING, WITH ASSISTANCE FROM THE SULFUR ANOMALOUS SIGNAL IN A HIGH-REDUNDANCY, 2.29 A WAVELENGTH DATA SET.
RMSD bond length	0.005
RMSD bond angle	1.300
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	CNX

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	36.120	2.020
High resolution limit [Å]	1.950	1.950
Rmerge	0.067	0.467
Number of reflections	81407
<I/σ(I)>	31.4	2.64
Completeness [%]	96.3	92
Redundancy	6.7	6.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	294	The purified protein was concentrated to 10-20 mg/ml and buffer exchanged into 25 mM Tris-HCl, pH 8.0, 30-50 mM NaCl, 25 mM sodium citrate. DMO crystals were grown using the vapor diffusion by sitting drop method, with a reservoir solution of 8-11% PEG 6000, 100 mM sodium acetate buffer pH 5.5, 1 M LiCl, and 4 uL droplets of varying protein-to-precipitant ratios., VAPOR DIFFUSION, SITTING DROP, temperature 294K