5ODC

Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus at 2.3 A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-04-14
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.73300
Spacegroup name	C 1 2 1
Unit cell lengths	378.047, 98.446, 137.884
Unit cell angles	90.00, 110.70, 90.00

Refinement procedure

Resolution	49.220 - 2.300
R-factor	0.1789
Rwork	0.178
R-free	0.20040
Structure solution method	SAD
RMSD bond length	0.005
RMSD bond angle	1.000
Data reduction software	XDS
Data scaling software	SCALA (3.3.22)
Phasing software	PHASER
Refinement software	BUSTER (2.10.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.220	2.420
High resolution limit [Å]	2.300	2.300
Rmerge	0.135	0.586
Rpim	0.053	0.282
Number of reflections	204479	25173
<I/σ(I)>	10.9	2.5
Completeness [%]	97.4	83
Redundancy	8.5	5.8
CC(1/2)	0.996	0.500

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	294	Crystallization was performed in an anaerobic chamber (95% N2/5% H2) with anoxic solution. The best-diffracting crystals were obtained using the sitting drop method in a crystallization plate (96-well 2-drop MRC Crystallization Plates in polystyrene, Molecular Dimensions, Suffolk, UK); the crystallization reservoir contained 100 mM Tris/HCl, pH 8.5, 30% (w/v) polyethylene glycol 4000, and 200 mM Na acetate trihydrate. Crystallization drop contained 1 ul HdrABC-MvhAGD at 25 mg/ml premixed with 2 mM FAD and 1 ul of precipitant. The crystals appear after 1-2 weeks and are cryoprotected in the crystallization solution supplemented with 30% glycerol.