9TN7

F420-nitrite reductase from Methanocaldococcus infernus at 1.58 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	2024-05-18
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.0332
Spacegroup name	I 2 2 2
Unit cell lengths	102.200, 114.110, 136.710
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.320 - 1.580
R-factor	0.1394
Rwork	0.138
R-free	0.16720
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.011
RMSD bond angle	1.314
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]	87.600	1.710
High resolution limit [Å]	1.580	1.580
Rmerge	0.124	1.957
Rmeas	0.129	2.032
Rpim	0.035	0.544
Number of reflections	89629	4481
<I/σ(I)>	11.4	1.5
Completeness [%]	94.8	52.6
Redundancy	13.5	13.8
CC(1/2)	0.998	0.558

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	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, 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 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 MES pH 6.5, and 400 mM Potassium chloride. The crystals appeared in 7 to 12 days.