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).