3GB4

Crystal Structure of Dicamba Monooxygenase with Non-heme Cobalt and Dicamba

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	2006-11-11
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.000
Spacegroup name	P 32
Unit cell lengths	81.550, 81.550, 161.290
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	20.000 - 2.050
R-factor	0.225
Rwork	0.221
R-free	0.26200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	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 the 2.29 A wavelength data set.
RMSD bond length	0.006
RMSD bond angle	1.320
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]	50.000	2.120
High resolution limit [Å]	2.050	2.050
Rmerge	0.068	0.485
<I/σ(I)>	23.1	1.66
Completeness [%]	99.6	99.9
Redundancy	4.1	4.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	294	Equal volumes of protein (10-20 mg/mL in 30 mM Tri-HCl,pH 8.0, 0.1 mM EDTA, with trace PMSF) and precipitant solution (15-20%(w/v) PEG 6000 and 0.1 M sodium citrate pH 6.0) were used to make the sitting drop. The data collection crystal was transferred to a cryo-amenable soak solution containing 24% PEG 6000, 24% glycerol, 0.1M HEPES-pH 7, 10 mM CoCl2, and 1.25 mM dicamba for 24 hours prior to being plunge-cooled in liquid nitrogen for data collection, VAPOR DIFFUSION, SITTING DROP, temperature 294K