8Q5V

MgADP-bound Fe protein of the molybdenum nitrogenase from Methanothermococcus thermolithotrophicus

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-01-28
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00004
Spacegroup name	P 1
Unit cell lengths	61.478, 87.594, 182.048
Unit cell angles	93.98, 98.58, 109.65

Refinement procedure

Resolution	39.930 - 2.740
R-factor	0.219
Rwork	0.217
R-free	0.25430
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	1.239
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]	71.000	2.900
High resolution limit [Å]	2.740	2.740
Rmerge	0.284	1.224
Rmeas	0.307	1.358
Rpim	0.117	0.509
Number of reflections	66142	3154
<I/σ(I)>	4.5	1.6
Completeness [%]	90.5	63.1
Redundancy	6.7	6.7
CC(1/2)	0.983	0.526

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293.15	The purified protein was crystallised at a final concentration of 32.7 mg/ml with 10 mM ATP and 10 mM MgCl2 by spotting 0.5 ul of crystallisation solution with 0.5 ul of protein sample. Crystallisation was done anaerobically in a Coy tent containing N2/H2 (97:3%) atmosphere. The screening was done at 20 degrees Celsius on 96-Well MRC 2-drop polystyrene (SWISSCI) plates containing 90 ul of crystallisation solution. The crystallisation solution in which crystals were obtained contained 20 % w/v polyethylene glycol 8000, 100 mM Tris pH 8.5 and 200 mM magnesium chloride. The crystals were soaked in the crystallisation solution supplemented with 30% glycerol prior freezing in liquid nitrogen.