4YX7
Complex of SpaO(SPOA1,2) 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 | 2013-11-22 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 1.075 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 62.092, 89.070, 62.092 |
Unit cell angles | 90.00, 114.94, 90.00 |
Refinement procedure
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 47.590 | 47.590 | 2.050 |
High resolution limit [Å] | 2.000 | 8.940 | 2.000 |
Rmerge | 0.102 | 0.062 | 0.530 |
Rpim | 0.050 | 0.030 | 0.253 |
Total number of observations | 210953 | 2212 | 15750 |
Number of reflections | 41204 | ||
<I/σ(I)> | 10.5 | 19.6 | 3.2 |
Completeness [%] | 99.5 | 92.8 | 99.8 |
Redundancy | 5.1 | 4.9 | 5.2 |
CC(1/2) | 0.994 | 0.991 | 0.828 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 293 | SpaO(145-213) + SpaO (232-297) + OrgB(1-30)::T4 lysozyme was concentrated to 18.5mg/mL 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 30% PEG3350, 10% glycerol, 200mM ammonium acetate, 100mM sodium acetate pH=5.0. |