5ODI

Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus cocrystallized with CoM-SH

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	370.528, 97.077, 135.149
Unit cell angles	90.00, 109.43, 90.00

Refinement procedure

Resolution	48.540 - 2.400
R-factor	0.2035
Rwork	0.202
R-free	0.24080
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5odc
RMSD bond length	0.006
RMSD bond angle	1.050
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]	48.540	2.530
High resolution limit [Å]	2.400	2.400
Rmerge	0.189	0.507
Rpim	0.071	0.188
Number of reflections	176807	25737
<I/σ(I)>	7.2	3.6
Completeness [%]	100.0	100
Redundancy	8	8.2
CC(1/2)	0.992	0.332

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 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, 1% (w/v) polyethylene glycol 3350, 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, 10 mM CoM-SH and 1 ul of precipitant. The crystals appear after 1-2 weeks and are cryoprotected in the crystallization solution supplemented with 30% glycerol