8RJA

Crystal structure of the F420-reducing formylmethanofuran dehydrogenase complex from the ethanotroph Candidatus Ethanoperedens thermophilum

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	2020-03-21
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00000
Spacegroup name	P 1 21 1
Unit cell lengths	107.627, 135.636, 149.902
Unit cell angles	90.00, 90.49, 90.00

Refinement procedure

Resolution	57.380 - 1.970
R-factor	0.1774
Rwork	0.176
R-free	0.21000
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.339
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]	57.380	2.187
High resolution limit [Å]	1.970	1.973
Rmerge	0.251	1.200
Rmeas	0.271	1.296
Rpim	0.101	0.486
Number of reflections	167948	8397
<I/σ(I)>	7.2	1.6
Completeness [%]	93.0	64.3
Redundancy	7	6.9
CC(1/2)	0.989	0.621

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	293.15	Crystals were obtained by initial screening at 20 degrees Celsius using the sitting drop method on a 96-Well MRC 2-Drop Crystallization Plates in polystyrene (SWISSCI). The crystallization reservoir contained 90 ul of mother liquor, the crystallization drop contained a mixture of 0.6 ul protein at 4.64 mg.ml-1 and 0.6 ul precipitant. The protein was in 25 mM Tris/HCl buffer pH 7.6, 10 % (v/v) glycerol, 2 mM DTT, 1 mM FAD and 1 mM FMN. The crystal was obtained in a Coy tent under an N2/H2 atmosphere (96:4 %). The crystallization reservoir contained 200 mM sodium acetate trihydrate, 100 mM sodium cacodylate, pH 6.5, and 18 % (w/v) polyethylene glycol 8,000. The crystal was soaked in the crystallization solution supplemented with 30 % (v/v) glycerol before freezing in liquid nitrogen.