3GOB
Crystal Structure of Dicamba Monooxygenase with Non-heme Cobalt and DCSA
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 | 81.010, 81.010, 161.050 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 20.000 - 2.050 |
R-factor | 0.252 |
Rwork | 0.232 |
R-free | 0.26700 |
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 a high-redundancy 2.29 A wavelength data set. |
RMSD bond length | 0.006 |
RMSD bond angle | 1.330 |
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 | 0.069 | 0.451 |
Number of reflections | 73352 | |
<I/σ(I)> | 22.8 | 1.91 |
Completeness [%] | 99.5 | 99.9 |
Redundancy | 4 | 4 |
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 Tris-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, 0.1 M HEPES pH 7.0, 10 mM CoCl2, and 1.25 mM DCSA for 24 hours prior to being plunge-cooled in liquid nitrogen for data collection. VAPOR DIFFUSION, SITTING DROP, temperature 294K |