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.