2FLQ

Crystal Structure of Nitric Oxide Synthase from Geobacillus Stearothermophilus (ATCC 12980) complexed with L-arginine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	100
Detector technology	CCD
Collection date	2005-03-12
Detector	ADSC QUANTUM 4
Wavelength(s)	1.002
Spacegroup name	P 21 21 2
Unit cell lengths	154.013, 118.721, 49.761
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 3.200
R-factor	0.229
Rwork	0.222
R-free	0.29500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Nitric Oxide Synthase from Bacillus Subtilis. (1M7V.pdb)
RMSD bond length	0.008
RMSD bond angle	1.511
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000
High resolution limit [Å]	3.200	3.200
Rmerge		0.313
Number of reflections	15805
<I/σ(I)>	12.9	5.5
Completeness [%]	92.0	92

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	298	GsNOS produced orthorhombic crystals of dimensions 200-400m in 24-48 h at 298K when grown by vapor diffusion from 45-50 mg/ml protein in 50mM Tris, 150mM NaCl, mixed with freshly dissolved 1-2 mM L-arginine and 1-2 mM L-tryptophan (Trp). The reservoir was mixed 1:1 with protein solution and contained 40-46% dioxane and 10% MPD, pH 7.5, VAPOR DIFFUSION, HANGING DROP