6CXK
E. coli DHFR substrate complex with Dihydrofolate
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 31-ID |
Synchrotron site | APS |
Beamline | 31-ID |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2017-11-30 |
Detector | RAYONIX MX-225 |
Wavelength(s) | 0.97931 |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 33.698, 51.525, 77.421 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 30.949 - 1.108 |
R-factor | 0.1792 |
Rwork | 0.179 |
R-free | 0.19610 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 7dfr |
RMSD bond length | 0.009 |
RMSD bond angle | 1.334 |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | PHASER |
Refinement software | PHENIX (1.8.4_1496) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 30.949 | 30.949 | 1.180 |
High resolution limit [Å] | 1.108 | 3.320 | 1.108 |
Rmerge | 0.067 | 0.024 | 0.861 |
Rmeas | 0.073 | 0.026 | 1.051 |
Number of reflections | 53265 | 2197 | 7579 |
<I/σ(I)> | 13.44 | 56.14 | 1.16 |
Completeness [%] | 97.9 | 99.1 | 87.3 |
Redundancy | 6.255 | 6.693 | 2.804 |
CC(1/2) | 0.999 | 0.999 | 0.571 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.5 | 293.15 | Sitting drop of final volume of 0.8 uL was set up by 1:1 v/v mixing of 15 mg/mL E. coli DHFR working stock with reservoir solution containing 0.1M MES pH 6.5, 27.5% PEG 3350, 0.4M MgCl2. E. coli DHFR working stock was prepared from a 1:1 v/v mixture of 30 mg/ml protein in 20 mM Tris pH 8, 1mM DTT with 50 mM HEPES pH 7.3, 100 mM NaCl. Crystals harvested after 2 weeks growth and cryoprotected with mitegen LV oil before flash freezing in N2. |