6V2A
Complex of double mutant (T89V,K162T) of E. coli L-asparaginase II with L-Asn
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | ROTATING ANODE |
Source details | RIGAKU MICROMAX-007 HF |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2018-03-15 |
Detector | DECTRIS EIGER X 4M |
Wavelength(s) | 1.5418 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 151.439, 62.594, 141.659 |
Unit cell angles | 90.00, 117.80, 90.00 |
Refinement procedure
Resolution | 25.980 - 2.000 |
R-factor | 0.1598 |
Rwork | 0.157 |
R-free | 0.20880 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 3eca |
RMSD bond length | 0.014 |
RMSD bond angle | 1.893 |
Data reduction software | HKL-2000 |
Data scaling software | HKL-2000 |
Phasing software | PHASER |
Refinement software | REFMAC (5.8.0253) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 40.000 | 40.000 | 2.030 |
High resolution limit [Å] | 2.000 | 5.420 | 2.000 |
Rmerge | 0.039 | 0.014 | 0.509 |
Rmeas | 0.048 | 0.016 | 0.633 |
Rpim | 0.027 | 0.009 | 0.371 |
Total number of observations | 207458 | ||
Number of reflections | 73175 | 4063 | 3421 |
<I/σ(I)> | 15.5 | ||
Completeness [%] | 92.6 | 99 | 86.7 |
Redundancy | 2.8 | 3.4 | 2.6 |
CC(1/2) | 0.999 | 0.690 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 4.7 | 293 | Crystals grown at 0.17 M sodium citrate (pH 6) and 17-18% (w/v) PEG3350. Soaked for 1-2 minutes (empirically determined for each crystal) in equivalent solution with 0.025% (w/v) glutaraldehyde. Finally transferred and soaked for 3-5 min in solution containing 40% (w/v) PEG3350, 5 mM L-Asn, and 0.17 M sodium citrate (pH 4.7) |