6YU9
CO-dehydrogenase homodimer from Clostridium autoethanogenum at 1.90-A resolution
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SLS BEAMLINE X06DA |
Synchrotron site | SLS |
Beamline | X06DA |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2019-12-14 |
Detector | DECTRIS PILATUS 2M-F |
Wavelength(s) | 1.00004 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 106.531, 109.446, 129.280 |
Unit cell angles | 90.00, 112.55, 90.00 |
Refinement procedure
Resolution | 49.750 - 1.904 |
R-factor | 0.1598 |
Rwork | 0.158 |
R-free | 0.20260 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 6ytt |
RMSD bond length | 0.011 |
RMSD bond angle | 1.120 |
Data reduction software | XDS |
Data scaling software | STARANISO |
Phasing software | MOLREP |
Refinement software | BUSTER (2.10.3 (19-MAR-2020)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 119.400 | 2.179 |
High resolution limit [Å] | 1.900 | 1.900 |
Rmerge | 0.205 | 0.804 |
Rmeas | 0.223 | 0.928 |
Rpim | 0.088 | 0.450 |
Number of reflections | 102786 | 5140 |
<I/σ(I)> | 7.7 | 1.8 |
Completeness [%] | 93.3 | 63.9 |
Redundancy | 6.2 | 4 |
CC(1/2) | 0.993 | 0.611 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.6 | 291.15 | Crystallization was performed in an anoxic chamber (N2/H2, 95:5 %) with anaerobic solutions. Crystals were obtained by initial screening at 291.15 K using the sitting drop method on a 96-well MRC 2 Crystallization Plates in polystyrene (SWISSCI). The crystallization reservoir contained 90 uL of mother liquor, crystallization drop contained a mixture of 0.6 uL protein and 0.6 uL precipitant. The protein concentration was 16.9 mg/mL in 25 mM Tris/HCl pH7.6, 10% glycerol (v/v) and 2 mM dithiothreitol. The best diffracting crystal of CODH was obtained by initial screening using the Shotgun (SG1) screen from Molecular dimensions. The crystallization reservoir contained 200 mM Magnesium acetate tetrahydrate; 20% (w/v) polyethylene glycol 3350. Crystals were cryo protected in the same solution supplemented with 25% glycerol (v/v). |