3RGW

Crystal structure at 1.5 A resolution of an H2-reduced, O2-tolerant hydrogenase from Ralstonia eutropha unmasks a novel iron-sulfur cluster

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2010-07-10
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.93950
Spacegroup name	P 21 21 21
Unit cell lengths	73.091, 95.646, 119.145
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	59.570 - 1.500
R-factor	0.14035
Rwork	0.140
R-free	0.15293
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1wul
RMSD bond length	0.010
RMSD bond angle	1.267
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHASER (& CNS (SIMULATED ANNEALING))
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	59.570	1.580
High resolution limit [Å]	1.500	1.500
Rmerge	0.088	0.434
Number of reflections	126968
<I/σ(I)>	11.2	3.7
Completeness [%]	100.0	100
Redundancy	6	5.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		282	20-30% polyethylene glycol 3350, 100 mM Bis-(2-hydroxy-ethyl)-amino-tris(hydroxymethyl)-methane buffer, pH 5.5-6.5, VAPOR DIFFUSION, SITTING DROP, temperature 282K