5ODR

Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with heterodisulfide for 2 minutes.

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	2016-06-23
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97916
Spacegroup name	C 1 2 1
Unit cell lengths	366.219, 96.920, 134.450
Unit cell angles	90.00, 108.30, 90.00

Refinement procedure

Resolution	90.260 - 2.200
R-factor	0.2048
Rwork	0.204
R-free	0.22480
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5odc
RMSD bond length	0.006
RMSD bond angle	0.900
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]	93.360	2.320
High resolution limit [Å]	2.200	2.200
Rmerge	0.181	0.897
Rpim	0.096	0.496
Number of reflections	224984	32472
<I/σ(I)>	4.6	1.3
Completeness [%]	99.4	98.7
Redundancy	4.3	4
CC(1/2)	0.983	0.563

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	294	All crystallization was performed in an anaerobic chamber (95% N2/5% H2) with anoxic solution using the sitting drop method (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. Crystal was soaked for 2 minutes in the crystallization solution supplemented with 66 mM of heterodisulfide (CoM-S-S-CoB). Then, the crystal was transferred in a solution containing the mother liquor with 30% glycerol and 10 mM CoM-S-S-CoB prior freezing.