4YX5

SpaO(SPOA1,2)

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, 0.979
Spacegroup name	P 41 21 2
Unit cell lengths	65.760, 65.760, 95.650
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.678 - 2.900
R-factor	0.2187
Rwork	0.212
R-free	0.27910
Structure solution method	SAD
RMSD bond length	0.010
RMSD bond angle	1.324
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]	54.190	54.190	3.080
High resolution limit [Å]	2.900	8.700	2.900
Rmerge	0.166	0.109	1.447
Rpim	0.034	0.024	0.283
Total number of observations	122494	4571	20277
Number of reflections	4975
<I/σ(I)>	14.4	26.9	2.7
Completeness [%]	99.3	95.2	99.3
Redundancy	24.6	21.2	26.2
CC(1/2)	0.996	0.995	0.856

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	SpaO(145-213) + SpaO (232-297) was concentrated to 12mg/mL and crystallized with 25% PEG400, 10% isopropanol, 100mM sodium citrate pH=5.6 at 277K. 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 mother liquor with the PEG400 concentration raised to 37.5%.