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). |