8P8G

Nitrogenase MoFe protein from A. vinelandii beta double mutant D353G/D357G

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	2021-10-16
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00002
Spacegroup name	C 1 2 1
Unit cell lengths	167.486, 74.469, 208.467
Unit cell angles	90.00, 103.25, 90.00

Refinement procedure

Resolution	48.320 - 1.550
R-factor	0.1566
Rwork	0.155
R-free	0.18720
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.012
RMSD bond angle	1.035
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]	101.460	1.760
High resolution limit [Å]	1.550	1.550
Rmerge	0.177	1.890
Rmeas	0.189	2.001
Rpim	0.066	0.652
Number of reflections	176043	8391
<I/σ(I)>	8.1	1.8
Completeness [%]	94.0	80.5
Redundancy	8.1	9.3
CC(1/2)	0.995	0.566

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.6	293.15	The beta-D353G/D357G MoFe-protein was crystallized anaerobically at 17.5 mg/mL under 100% N2 atmosphere. The protein was spotted as a sitting drop to 96-Well MRC 2-Drop polystyrene Crystallization Plates (SWISSCI) containing 90 uL of crystallization solution in the reservoir. Each drop contained 0.5 uL of protein sample and 0.5 uL of crystallization solution. Crystals were obtained in the crystallization solution containing 10 % w/v Polyethylene glycol 10,000; 2 % v/v 1,4-Dioxane; 100 mM tri-Sodium citrate; pH 5.6, and 1 mM polyoxotungstate [TeW6O24]6- (TEW). Sealed plates were stored inside a Coy anaerobic chamber filled with an atmosphere of N2:H2 97:3 at 20 C. Crystals were soaked in the crystallization solution supplemented with 30% v/v ethylene glycol for a few seconds before freezing in liquid nitrogen.