5DEN

The First Structure of a Full-Length Mammalian Phenylalanine Hydroxylase Reveals the Architecture of an Auto-inhibited Tetramer

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-09-25
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.1
Spacegroup name	C 1 2 1
Unit cell lengths	113.780, 89.160, 196.810
Unit cell angles	90.00, 104.53, 90.00

Refinement procedure

Resolution	38.100 - 2.900
R-factor	0.2417
Rwork	0.238
R-free	0.30310
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phz
RMSD bond length	0.002
RMSD bond angle	0.381
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.10pre_2120: ???)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.100	3.004
High resolution limit [Å]	2.900	2.900
Rmerge	0.068	0.373
Number of reflections	38661
<I/σ(I)>	9.06	1.56
Completeness [%]	91.0	76.49
Redundancy	1.9	1.56

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293.15	Protein was stored at -80C, then thawed in a water bath at 25C for ~ 20 minutes. The protein, at a concentration of 9.5 mg/mL was diluted to 5.5 mg/mL using 30 mM Tris pH 7.4, 116 mM KCl, 15% glycerol. The diluted protein was left at 4C for ~ hours, then at room temperature for ~ 20 minutes prior to preparation of the crystallization tray. The 2 uL hanging drop (containing 1:1 protein:reservoir) hung over a reservoir solution containing 140 mM Na-acetate, 70 mM Na-citrate, 100 mM Na-cacodylate (pH 6.5), and 31.5% PEG 1,000 (Hampton Research).