4U9G

Crystal structure of an H-NOX protein from S. oneidensis in the Fe(II)CO 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)	1.00
Spacegroup name	P 63 2 2
Unit cell lengths	164.173, 164.173, 102.366
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	43.010 - 2.250
R-factor	0.182
Rwork	0.182
R-free	0.19300
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4u99
RMSD bond length	0.009
RMSD bond angle	0.887
Refinement software	PHENIX ((PHENIX.REFINE: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.007	2.320
High resolution limit [Å]	2.250	2.250
Rmerge	0.028	0.446
Number of reflections	70856
<I/σ(I)>	20.2	1.8
Completeness [%]	96.7	98.1
Redundancy	11.1	11.1

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, CRYSTALS WERE TRANSFERRED UNDER A LAYER OF OIL AND INCUBATED WITH CARBON MONOXIDE-SATURATED CRYOPROTECTANT FOR 15-45 MINUTES. CRYSTAL GROWTH AND MANIPULATION WAS PERFORMED ANAEROBICALLY.