4U99

Crystal structure of an H-NOX protein from S. oneidensis in the Fe(II) ligation state, Q154A/Q155A/K156A mutant

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	CCD
Collection date	2014-01-09
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.000
Spacegroup name	P 63 2 2
Unit cell lengths	164.003, 164.003, 101.718
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	47.880 - 2.000
R-factor	0.173
Rwork	0.172
R-free	0.19100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3tf8
RMSD bond length	0.005
RMSD bond angle	0.865
Refinement software	PHENIX ((PHENIX.REFINE: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.882	2.050
High resolution limit [Å]	2.000	2.000
Rmerge	0.021	0.618
Number of reflections	102223
<I/σ(I)>	0.208	1.4
Completeness [%]	98.8	97.9
Redundancy	7.6	6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.3	298	Obtained by equilibrating a 2 uL drop of 1:1 protein:reservoir against a 700 uL reservoir containing 1.6-1.9 M DL-malic acid (pH 7.3). For cryoprotection, 2 uL of mother liquor containing 10% glycerol was added directly to the drop and crystals were serial transferred into mother liquor solution containing 5, 7.5 and 10% glycerol prior to flash freezing in liquid nitrogen. Crystal growth and manipulation was performed anaerobically.