5KR5
Directed Evolution of Transaminases By Ancestral Reconstruction. Using Old Proteins for New Chemistries
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | AUSTRALIAN SYNCHROTRON BEAMLINE MX2 |
Synchrotron site | Australian Synchrotron |
Beamline | MX2 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2014-03-01 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 0.95370 |
Spacegroup name | C 2 2 21 |
Unit cell lengths | 98.451, 102.538, 199.781 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 48.600 - 2.100 |
R-factor | 0.20597 |
Rwork | 0.204 |
R-free | 0.23987 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5kqt |
RMSD bond length | 0.009 |
RMSD bond angle | 1.424 |
Data reduction software | XDS |
Data scaling software | Aimless |
Phasing software | PHASER |
Refinement software | REFMAC (5.8.0155) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 48.600 | 2.150 |
High resolution limit [Å] | 2.100 | 2.100 |
Rmerge | 0.170 | 0.899 |
Number of reflections | 59261 | |
<I/σ(I)> | 9.5 | 2.2 |
Completeness [%] | 99.6 | 97.6 |
Redundancy | 7.3 | 7.3 |
CC(1/2) | 0.994 | 0.670 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 293 | 150 plus 150 nL drops with protein at 4 mg/mL and reservoir conditions of 15% PEG 8000, 11 mM calcium acetate, 100 mM Tris pH 7.1. Microcrystals were used as nucleating agents to obtain full size crystals. |