8Q5T

Nitrogenase Fe protein from Methanothermococcus thermolithotrophicus, monoclinic crystalline form at 2.31-A resolution

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	2019-11-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.74013
Spacegroup name	P 1 21 1
Unit cell lengths	52.821, 83.984, 116.598
Unit cell angles	90.00, 91.14, 90.00

Refinement procedure

Resolution	48.470 - 2.310
R-factor	0.1963
Rwork	0.194
R-free	0.23490
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.874
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]	68.140	2.350
High resolution limit [Å]	2.310	2.310
Rmerge	0.240	1.342
Rmeas	0.255	1.460
Rpim	0.085	0.566
Number of reflections	45087	2223
<I/σ(I)>	5.2	1.5
Completeness [%]	100.0	100
Redundancy	8.7	6.5
CC(1/2)	0.990	0.517

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.6	293.15	The purified protein was crystallised at a final concentration of 6 mg/ml by spotting 0.5 ul of crystallisation solution with 0.5 ul of protein sample. Crystallisation was done anaerobically in a Coy tent containing a 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 30 % v/v 2-methyl-2,4-pentanediol, 100 mM sodium acetate, pH 4.6 and 20 mM calcium chloride.