4YX1

SpaO(SPOA2)

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	2013-07-31
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.979
Spacegroup name	P 1 21 1
Unit cell lengths	35.000, 41.270, 48.000
Unit cell angles	90.00, 103.92, 90.00

Refinement procedure

Resolution	31.262 - 1.350
R-factor	0.1747
Rwork	0.172
R-free	0.20530
Structure solution method	SAD
RMSD bond length	0.007
RMSD bond angle	1.093
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]	41.270	41.270	1.370
High resolution limit [Å]	1.350	7.390	1.350
Rmerge	0.146	0.122	1.281
Rpim	0.059	0.054	0.511
Total number of observations	203967	849	10254
Number of reflections	29266
<I/σ(I)>	8.6	15.5	2.1
Completeness [%]	99.7	76.7	100
Redundancy	7	5.6	7.1
CC(1/2)	0.994	0.981	0.750

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	SpaO(232-297) was concentrated to 8mg/mL and crystallized with 35% PEG400, 200mM calcium acetate, 100mM sodium acetate pH=5.0. Crystals were cryoprotected in the mother liquor. 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.