4YXA
Complex of SpaO(SPOA1,2 SeMet) and OrgB(APAR)::T4lysozyme fusion protein
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X29A |
Synchrotron site | NSLS |
Beamline | X29A |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2014-03-12 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 0.979 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 62.880, 88.500, 63.320 |
Unit cell angles | 90.00, 116.07, 90.00 |
Refinement procedure
Resolution | 45.804 - 2.350 |
R-factor | 0.2027 |
Rwork | 0.198 |
R-free | 0.26180 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 4yx7 |
RMSD bond length | 0.011 |
RMSD bond angle | 1.464 |
Data scaling software | Aimless (0.2.7) |
Phasing software | PHENIX |
Refinement software | PHENIX |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 47.610 | 47.610 | 2.430 |
High resolution limit [Å] | 2.350 | 9.100 | 2.350 |
Rmerge | 0.088 | 0.056 | 0.617 |
Rpim | 0.037 | 0.023 | 0.256 |
Total number of observations | 169948 | 2593 | 16809 |
Number of reflections | 25759 | ||
<I/σ(I)> | 12.8 | 29.5 | 2.6 |
Completeness [%] | 99.0 | 86.5 | 99.6 |
Redundancy | 6.6 | 6.4 | 6.6 |
CC(1/2) | 0.997 | 0.997 | 0.816 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 293 | SpaO(145-213, SeMet) + SpaO (232-297, SeMet) + OrgB(1-30)::T4 lysozyme (native) was concentrated to 18mg/mL, supplemented with 50mM maltose, and crystallized with 25% PEG3350, 200mM ammonium formate, 100mM sodium acetate pH=5.0. Microseeding was employed to enhance crystal uniformity and diffraction. Briefly, crystals to be seeded were harvested in precipitant solution and vortexed in a microfuge tube with a small stir bar for ~60 seconds. The slurry of microseeds was serially dilluted (5-10-fold steps) in precipitant solution and 5 selected microseed-precipitant mixtures were mixed with fresh protein as in a normal hanging drop experiment. Crystals were cryoprotected in 25% PEG3350, 10% ethylene glycol, 200mM ammonium formate, 100mM sodium acetate pH=5.0, 50mM maltose. |