4U9K

Crystal structure of an H-NOX protein from S. oneidensis in the Mn(II)NO 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	2013-12-17
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.00
Spacegroup name	P 63 2 2
Unit cell lengths	164.303, 164.303, 101.679
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	47.875 - 2.450
R-factor	0.1835
Rwork	0.182
R-free	0.20690
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4u9j
RMSD bond length	0.022
RMSD bond angle	1.643
Refinement software	PHENIX ((phenix.refine: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.875	2.550
High resolution limit [Å]	2.450	2.450
Rmerge	0.032	0.663
Number of reflections	56342
<I/σ(I)>	19.3	1.6
Completeness [%]	99.8	100
Redundancy	10.7	11

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 ANAEROBIC CRYOPROTECTANT CONTAINING APPROXIMATELY 0.001 M NITRIC OXIDE FOR 15-45 MINUTES. CRYSTAL GROWTH AND MANIPULATION WAS PERFORMED ANAEROBICALLY.