5V1S
Crystal structure of Streptococcus suis SuiB bound to S-adenosylmethionine
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 23-ID-D |
Synchrotron site | APS |
Beamline | 23-ID-D |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2016-02-19 |
Detector | DECTRIS PILATUS3 6M |
Wavelength(s) | 1.0332 |
Spacegroup name | P 21 21 2 |
Unit cell lengths | 114.787, 85.473, 109.864 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 29.470 - 2.492 |
R-factor | 0.2172 |
Rwork | 0.215 |
R-free | 0.26970 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5v1q |
RMSD bond length | 0.008 |
RMSD bond angle | 0.799 |
Data reduction software | XDS |
Data scaling software | XDS |
Phasing software | PHENIX |
Refinement software | PHENIX (1.9_1692) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 29.470 | 2.590 |
High resolution limit [Å] | 2.490 | 2.490 |
Rmerge | 0.067 | 0.839 |
Number of reflections | 38407 | |
<I/σ(I)> | 17.7 | 2 |
Completeness [%] | 99.8 | 99 |
Redundancy | 6.5 | 5.9 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 285 | A solution containing 18.9 mg/mL His6-tagged SuiB in in storage buffer [100 mM HEPES, pH 7.5, 300 mM KCl, 5 mM DTT, 10% (v/v) glycerol] was mixed 1:1 with precipitant solution. Small clusters of crystals were formed within 24 hrs. A seed stock was then produced by combining a single sitting well with 10 uL of reservoir solution and 10 uL of enzyme at 8 mg/mL, followed by brief vortexing. Following seeding, a resulting crystal was incubated in precipitant solution containing ~6 mM SAM for 30 min prior to transfer to cryosolution.The precipitant solution was 100 mM MES, pH 6.0, 15% (w/v) PEG 3350. |