9R5I

Dimeric state of the F420-reducing hydrogenase from Methanothermococcus thermolithotrophicus in crystalline form 3

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-04-28
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.73891
Spacegroup name	P 31 2 1
Unit cell lengths	196.606, 196.606, 192.072
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	77.830 - 3.130
R-factor	0.176
Rwork	0.174
R-free	0.20590
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.935
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]	127.410	3.350
High resolution limit [Å]	3.130	3.130
Rmerge	0.304	2.449
Rmeas	0.312	2.510
Rpim	0.069	0.545
Number of reflections	52281	2616
<I/σ(I)>	10	1.6
Completeness [%]	95.1	67.7
Redundancy	20.2	21.1
CC(1/2)	0.997	0.620

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	293.15	The enzyme was crystallized at a concentration of 38.8 mg.ml-1 in 25 mM Tris/HCl pH 7.6, 10% (v/v) glycerol, and 2 mM dithiothreitol. Crystals were obtained anaerobically in a Coy tent filled with a N2/H2 (97:3 %) atmosphere by initial screening at 20 degrees Celsius using the sitting-drop method on 96-well MRC two-drop crystallisation plates in polystyrene (SWISSCI). The reservoir contained 90 ul of the following crystallization solution: 30 % (v/v) Polyethylene glycol 400, 100 mM MES pH 6.5, and 200 mM LiSO4. 0.7 ul of protein and 0.7 ul of the crystallization solution were spotted.