3MIG

Oxidized (Cu2+) peptidylglycine alpha-hydroxylating monooxygenase (PHM) with bound nitrite, obtained in the presence of substrate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X4C
Synchrotron site	NSLS
Beamline	X4C
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2007-11-16
Detector	MAR scanner 345 mm plate
Wavelength(s)	0.98
Spacegroup name	P 21 21 21
Unit cell lengths	68.356, 68.783, 79.979
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	52.000 - 2.700
R-factor	0.22387
Rwork	0.221
R-free	0.27236
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phm
RMSD bond length	0.012
RMSD bond angle	1.221
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	52.000	2.800
High resolution limit [Å]	2.700	2.700
Number of reflections	10776
<I/σ(I)>	35.7	2.1
Completeness [%]	99.0	91.4
Redundancy	7	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	293	Crystallization: 0.1-0.5mM CuSO4, 1.25mM NiCl2, 100mM sodium cacodylate pH=5.5, 3mM sodium azide and 5% glycerol. The crystal was first soaked in 1mM substrate (Ac-3,5-diI-YG) for 1 hour and then soaked in 300mM NaNO2/1mM substrate for 14 hours at 293K., VAPOR DIFFUSION, HANGING DROP