9R52

Dimeric state of the F420-reducing hydrogenase from Methanothermococcus thermolithotrophicus in crystalline form 2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06SA
Synchrotron site	SLS
Beamline	X06SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-12-03
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.00003
Spacegroup name	C 2 2 21
Unit cell lengths	95.028, 163.482, 128.826
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	64.410 - 1.650
R-factor	0.1432
Rwork	0.141
R-free	0.18260
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.189
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHASER
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	69.270	1.790
High resolution limit [Å]	1.650	1.650
Rmerge	0.112	2.354
Rmeas	0.115	2.421
Rpim	0.026	0.560
Number of reflections	92187	4460
<I/σ(I)>	16.1	1.5
Completeness [%]	94.1	59.9
Redundancy	20.1	18.4
CC(1/2)	0.999	0.588

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	Crystals were obtained anaerobically in a Coy tent filled with a N2/H2 (97:3%) atmosphere at 20 degree Celsius using the sitting/drop method in CombiClover crystallisation plates. The enzyme was used at a concentration of 6.1 mg/ml in 25 mM Tris/HCl pH 7.6, 10% (v/v) glycerol, 2 mM dithiothreitol and 2 mM flavin adenine dinucleotide (FAD). 2 ul of the protein sample was mixed with 2 ul of the following crystallisation solution made of 45 % (w/v) Pentaerythritol propoxylate (17/8 PO/OH) and 100 mM Tris pH 8.5.