5ODQ

Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with bromoethanesulfonate.

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)	0.91941
Spacegroup name	C 1 2 1
Unit cell lengths	366.325, 97.352, 133.276
Unit cell angles	90.00, 108.37, 90.00

Refinement procedure

Resolution	48.680 - 2.150
R-factor	0.1973
Rwork	0.196
R-free	0.21860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5odc
RMSD bond length	0.006
RMSD bond angle	0.940
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]	48.850	2.270
High resolution limit [Å]	2.150	2.150
Rmerge	0.210	0.606
Rpim	0.086	0.249
Number of reflections	241487	35106
<I/σ(I)>	8	3.3
Completeness [%]	100.0	100
Redundancy	6.9	6.8
CC(1/2)	0.991	0.327

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. The soaking experiment with the bromoethanesulfonate was done by soaking the crystals in the same crystallization solution supplemented with 66 mM of bromoethanesulfonate for 7 min. The crystal has been cryo-protected using the the mother liquor solution supplemented with 30% glycerol and 10 mM bromoethanesulfonate.