3IEC
Helicobacter pylori CagA Inhibits PAR1/MARK Family Kinases by Mimicking Host Substrates
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X29A |
Synchrotron site | NSLS |
Beamline | X29A |
Temperature [K] | 103 |
Detector technology | CCD |
Collection date | 2008-08-08 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 1.0809 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 93.466, 93.256, 113.955 |
Unit cell angles | 90.00, 100.94, 90.00 |
Refinement procedure
Resolution | 40.960 - 2.200 |
R-factor | 0.206 |
Rwork | 0.204 |
R-free | 0.24800 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.015 |
RMSD bond angle | 1.507 |
Data reduction software | DENZO |
Data scaling software | SCALEPACK |
Phasing software | PHASER |
Refinement software | REFMAC |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 50.000 | 50.000 | 2.280 |
High resolution limit [Å] | 2.200 | 4.740 | 2.200 |
Rmerge | 0.089 | 0.048 | 0.706 |
Number of reflections | 97843 | ||
<I/σ(I)> | 10.5 | ||
Completeness [%] | 99.9 | 99.1 | 100 |
Redundancy | 6.2 | 5.9 | 6.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 296 | Initial crystals were grown by vapor diffusion using hanging drops formed from mixing a 2:1 volume ratio of the protein complex with an equilibration buffer consisting of 22.5-25% polyethylene glycol (PEG) molecular weight 3350 Da, 300 mM Li2SO4, and 100 mM Bis-Tris pH 5.8-6.2, at 23 C. Higher quality crystals were obtained by micro-seeding and addition of 5% PEG 400 as an additive. For cryoprotection crystals where transferred directly into a buffer with a 25% PEG 3350 Da, 5-10% PEG 400 Da, 300mM Li2SO4, 100mM Bis-Tris pH 5.8-6.2, 7.5% glycerol, and flash-cooled immediately afterward to -160 C. , VAPOR DIFFUSION, HANGING DROP, temperature 296K |