2YCN
Y71F mutant of tyrosine phenol-lyase from Citrobacter freundii in complex with quinonoid intermediate formed with 3-fluoro-L-tyrosine
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE BM14 |
Synchrotron site | ESRF |
Beamline | BM14 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2002-05-04 |
Detector | MARMOSAIC 225 mm CCD |
Spacegroup name | P 21 21 2 |
Unit cell lengths | 133.635, 144.397, 59.660 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 98.060 - 2.040 |
R-factor | 0.16405 |
Rwork | 0.163 |
R-free | 0.20886 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 2ez2 |
RMSD bond length | 0.014 |
RMSD bond angle | 1.383 |
Data reduction software | DENZO |
Data scaling software | SCALEPACK |
Phasing software | AMoRE |
Refinement software | REFMAC (5.5.0109) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 17.000 | 2.090 |
High resolution limit [Å] | 2.040 | 2.040 |
Rmerge | 0.050 | 0.350 |
Number of reflections | 71319 | |
<I/σ(I)> | 13.7 | 3.2 |
Completeness [%] | 96.0 | 84 |
Redundancy | 3 | 1.9 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 8 | 293 | CRYSTALS WERE GROWN AT 277 AND 293 K USING THE HANGING DROP VAPOR DIFFUSION METHOD. THE BEST CRYSTALS WERE OBTAINED BY MIXING 2 UL OF THE PROTEIN SOLUTION (18-20 MG/ML) CONTAINING 50 MM K-PHOSPHATE PH 8.0, 0.5 MM PLP, 1 MM DDT WITH AN EQUAL VOLUME OF THE RESERVOIR SOLUTION CONTAINING 50 MM TRIETHANOLAMINE BUFFER (PH 8.0), 0.5 MM PLP, 2 MM DDT, 0.4 M KCL, AND 35-38% (W/V) POLY(ETHYLENE GLYCOL) 5000 MONOMETHYL ETHER. COMPLEX OF Y71F TPL WITH 3-FLUORO-L-TYROSINE WAS PREPARED BY SOAKING CRYSTALS IN THE STABILIZING SOLUTION CONTAINING 10 MM 3- FLUORO-L-TYROSINE FOR ABOUT 30 SECONDS. |