6V2M
Structure of Escherichia coli Asp269Asn mutant phosphoenolpyruvate carboxykinase
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 | 124.780, 94.220, 46.540 |
Unit cell angles | 90.00, 94.79, 90.00 |
Refinement procedure
Resolution | 46.380 - 1.660 |
Rwork | 0.185 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1ayl |
Data reduction software | XDS |
Data scaling software | XDS |
Phasing software | MOLREP |
Refinement software | PHENIX (1.17.1_3660) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 47.110 | 1.719 |
High resolution limit [Å] | 1.660 | 1.660 |
Rmerge | 0.115 | 2.915 |
Rmeas | 0.132 | 3.340 |
Rpim | 0.064 | 1.613 |
Number of reflections | 63107 | 6242 |
<I/σ(I)> | 7.96 | 0.65 |
Completeness [%] | 99.7 | 98.71 |
Redundancy | 4.2 | 4.2 |
CC(1/2) | 0.997 | 0.175 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | MICROBATCH | 293 | 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 |