5LTE

Crystal structure of the alpha subunit of heme dependent oxidative N-demethylase (HODM)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-04-26
Detector	DECTRIS PILATUS3 X 2M
Wavelength(s)	0.917
Spacegroup name	P 43 21 2
Unit cell lengths	79.908, 79.908, 144.712
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	26.820 - 1.650
R-factor	0.16493
Rwork	0.163
R-free	0.19359
Structure solution method	MIR
RMSD bond length	0.024
RMSD bond angle	2.230
Data reduction software	XDS
Data scaling software	XDS
Phasing software	MLPHARE
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	26.820	1.700
High resolution limit [Å]	1.650	1.650
Rmerge	0.061	0.783
Number of reflections	53715
<I/σ(I)>	15.4	3.8
Completeness [%]	99.9	100
Redundancy	12.1	12.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277	The HODM-heme subunit was crystallised at 3 mg ml-1 in 50 mM KPi, 100 mM KCl, pH 7.5. Initial crystallisation conditions were identified using the JCSG+ matrix screen (Molecular dimensions). Crystals suitable for diffraction experiments were obtained by sitting drop vapour diffusion at 277 K in 400 nL drops containing equal volumes of protein and a solution containing 30% PEG 2K MME and 0.1 M potassium thiocyanate