5LTH

Crystal structure of the alpha subunit of heme dependent oxidative N-demethylase (HODM) in complex with the dimethylamine substrate

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 6M
Wavelength(s)	0.917
Spacegroup name	P 43 21 2
Unit cell lengths	80.380, 80.380, 144.790
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.010 - 1.760
R-factor	0.16626
Rwork	0.165
R-free	0.18882
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	5lte
RMSD bond length	0.027
RMSD bond angle	2.318
Data reduction software	XDS
Data scaling software	XDS
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	33.010	1.810
High resolution limit [Å]	1.760	1.760
Rmerge	0.037	0.752
Number of reflections	45331
<I/σ(I)>	18.2	3.5
Completeness [%]	99.9	100
Redundancy	12.9	12.3

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.