6GGU

Crystal structure of native FE-hydrogenase from Methanothermobacter marburgensis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM30A
Synchrotron site	ESRF
Beamline	BM30A
Temperature [K]	100
Detector technology	CCD
Collection date	2016-09-21
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97980
Spacegroup name	P 63 2 2
Unit cell lengths	144.060, 144.060, 95.060
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.840 - 2.600
R-factor	0.191
Rwork	0.188
R-free	0.23500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4jjf
RMSD bond length	0.008
RMSD bond angle	1.030
Data reduction software	XDS
Data scaling software	SCALA (3.3.22)
Phasing software	PHASER
Refinement software	BUSTER (2.10.3)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.530	2.740
High resolution limit [Å]	2.600	2.600
Rmerge	0.311	0.914
Rmeas	0.323
Rpim	0.085	0.244
Number of reflections	18373	2608
<I/σ(I)>	9.2	3.4
Completeness [%]	99.9	99.7
Redundancy	14.6	15
CC(1/2)	0.989	0.441

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	293	[Fe]-hydrogenase was crystallized under red light condition, in an anaerobic tent containing a gas phase of 100% N2 by the sitting drop vapuor diffusion method. The crystallization drops contained 0.7 ul of 30 mg.ml-1 protein and 2 mM methenyl-H4MPT+ mixed with 0.7 ul of 0.1 M MES pH 6.0 and 0.8 M ammonium sulfate and spotted on 96-well 2-drop MRC crystallization plates in polystyrene (Molecular Dimensions, Suffolk, UK). The best crystal appeared within 2.5 months.