9TN9

F420-nitrite reductase from Methanocaldococcus infernus soaked with 50 mM nitrite

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	2024-05-18
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.03320
Spacegroup name	I 2 2 2
Unit cell lengths	101.760, 113.000, 135.800
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.020 - 1.200
R-factor	0.117
Rwork	0.116
R-free	0.14270
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.013
RMSD bond angle	1.481
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHENIX
Refinement software	PHENIX ((1.21.2_5419: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	86.860	1.326
High resolution limit [Å]	1.200	1.205
Rmerge	0.067	0.938
Rmeas	0.074	1.074
Rpim	0.031	0.513
Number of reflections	175298	8765
<I/σ(I)>	11.9	1.6
Completeness [%]	95.1	56.9
Redundancy	5.4	4.2
CC(1/2)	0.998	0.470

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.6	293.15	Prior to crystallisation, fresh sample was centrifuged at 13 000 g for 3 min to remove macro-aggregates and dust. The sample was crystallised in an anaerobic chamber filled with a N2/H2 (97:3%) atmosphere, at 20 degrees Celsius. The crystallisation was done in 96-Well MRC 2-Drop polystyrene plates (SWISSCI) containing 90 uL of crystallisation solution in the reservoir. 0.7 uL of enzyme at 6.5 mg/ml + 1 mM FAD was mixed with 0.7 uL of the crystallisation solution. The crystallisation solution contained 45 % w/v pentaerythritol propoxylate (5/4 PO/OH), 100 mM sodium acetate pH 4.6, and 400 mM potassium chloride. Prior to flash-freezing in liquid nitrogen, crystals were soaked in the crystallisation solution supplemented with 50 mM sodium nitrite for 5 min.