3ZEA

3D structure of the NiFeSe hydrogenase from D. vulgaris Hildenborough in the reduced state at 1.82 Angstroms

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2011-11-21
Detector	DECTRIS PILATUS 6M
Spacegroup name	P 31 2 1
Unit cell lengths	61.796, 61.796, 339.297
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	56.550 - 1.820
R-factor	0.1249
Rwork	0.124
R-free	0.14680
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2wpn
RMSD bond length	0.010
RMSD bond angle	1.251
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	56.600	1.930
High resolution limit [Å]	1.820	1.820
Rmerge	0.040	0.080
Number of reflections	67904
<I/σ(I)>	23.3	9.3
Completeness [%]	97.7	89.9
Redundancy	4.1	3.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.1		CRYSTALS WERE OBTAINED USING THE SITTING-DROP VAPOR DIFFUSION METHOD. 1 UL OF A RESERVOIR SOLUTION CONTAINING 16% PEG 8000 (W/V) AND 0.05 M KH2PO4 PH 4.1 WAS MIXED WITH AN EQUAL VOLUME OF A SOLUTION COMPOSED OF 11 MG/ML PROTEIN IN 20 MM TRIS-HCL BUFFER PH 7.6, AND EQUILIBRATED AGAINST A 500 UL RESERVOIR.