6COM
2.3A crystal structure of E. coli phosphoenolpyruvate carboxykinase mutant Asp269Asn
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | CLSI BEAMLINE 08B1-1 |
Synchrotron site | CLSI |
Beamline | 08B1-1 |
Temperature [K] | 105 |
Detector technology | CCD |
Collection date | 2014-02-28 |
Detector | RAYONIX MX300HE |
Wavelength(s) | 0.98 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 124.595, 94.175, 46.457 |
Unit cell angles | 90.00, 94.77, 90.00 |
Refinement procedure
Resolution | 47.087 - 2.300 |
R-factor | 0.1535 |
Rwork | 0.148 |
R-free | 0.20920 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1ayl |
RMSD bond length | 0.008 |
RMSD bond angle | 0.963 |
Data reduction software | HKL-2000 |
Data scaling software | d*TREK |
Phasing software | PHASER |
Refinement software | PHENIX (dev_2398) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 50.000 | 2.190 |
High resolution limit [Å] | 2.110 | 2.110 |
Rmerge | 0.250 | |
Number of reflections | 77150 | |
<I/σ(I)> | 5.8 | |
Completeness [%] | 99.8 | |
Redundancy | 3.5 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | MICROBATCH | 8.2 | 293 | A 2ul drop with 2 mg/ml protein, 5 mM Calcium chloride, 5mM Magnesium chloride, 2 mM ATP, 2mM Pyruvate, 1 mM EDTA, 200 mM Ammonium acetate, 100 mM Sodium acetate, 0.01 mM DTT, 30% PEG 4000 was added to 2 ul drop containing 2 M sodium aceate, 0.1 M Tris pH 8.2 30% PEG 4000. After a week a rod like crystal was removed and soaked in a solution with 30% glycerol 1mM EDTA, 100 mM sodium acetate 200mM ammonium acetate and 12% PEG 4000. The crystal was put into a loop and flash cooled in liquid notrogen |