8RIU

Crystal structure of the F420-reducing carbon monoxide dehydrogenase component from the ethanotroph Candidatus Ethanoperedens thermophilum

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-11-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.97856
Spacegroup name	P 21 21 21
Unit cell lengths	97.070, 159.213, 191.444
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.800 - 1.890
R-factor	0.1662
Rwork	0.165
R-free	0.18730
Structure solution method	SAD
RMSD bond length	0.009
RMSD bond angle	1.233
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHENIX
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	122.410	2.100
High resolution limit [Å]	1.890	1.890
Rmerge	0.149	1.583
Rmeas	0.155	1.650
Rpim	0.042	0.460
Number of reflections	164775	8240
<I/σ(I)>	10.8	1.7
Completeness [%]	95.9	73.6
Redundancy	13.6	12.2
CC(1/2)	0.999	0.639

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6	293.15	Crystallisation was performed anaerobically at 20 degree Celsius using the sitting drop method on 96-Well MRC 2-Drop polystyrene Crystallisation Plates (SWISSCI) in a Coy tent containing a N2/H2 (97:3%) atmosphere. The reservoir chamber was filled with 90 ul of crystallisation condition and the drop was formed by spotting 0.6 ul protein with 0.6 ul of 20% (w/v) PEG 6,000; 100 mM MES pH6 and 200 mM Ammonium chloride. The protein was crystallized at 13.0 mg/ml in 25 mM Tris/HCl pH 7.6, 2 mM dithiothreitol, 1 mM flavin adenine dinucleotide and 1 mM Flavin mononuleotide and 10% (v/v) glycerol. Crystals were cryo-protected by soaking in 20% (w/v) PEG 6,000; 100 mM MES pH6 and 200 mM Ammonium chloride supplemented with 30% v/v glycerol for a few seconds.