4U9J

Crystal structure of an H-NOX protein from S. oneidensis in the Mn(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	2013-09-18
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.00
Spacegroup name	P 63 2 2
Unit cell lengths	163.801, 163.801, 102.082
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	48.027 - 2.100
R-factor	0.1723
Rwork	0.171
R-free	0.19720
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4u99
RMSD bond length	0.021
RMSD bond angle	1.650
Refinement software	PHENIX ((phenix.refine: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.027	2.160
High resolution limit [Å]	2.100	2.100
Rmerge	0.032	0.655
Number of reflections	86341
<I/σ(I)>	12.8	1.1
Completeness [%]	96.5	98.6
Redundancy	4.8	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.3	293	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.