3O65

Crystal structure of a Josephin-ubiquitin complex: Evolutionary restraints on ataxin-3 deubiquitinating activity

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X6A
Synchrotron site	NSLS
Beamline	X6A
Detector technology	CCD
Collection date	2008-10-10
Detector	ADSC QUANTUM 210
Wavelength(s)	0.9780
Spacegroup name	P 3 2 1
Unit cell lengths	159.150, 159.150, 146.290
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.903 - 2.700
R-factor	0.1781
Rwork	0.176
R-free	0.22420
Structure solution method	SAD
RMSD bond length	0.009
RMSD bond angle	1.140
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	RESOLVE (2.14)
Refinement software	PHENIX (1.6.1_357)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]			2.770
High resolution limit [Å]	2.700	12.080	2.700
Rmerge	0.096	0.040	0.588
Number of reflections	112055	992	7798
<I/σ(I)>	15.42	37.3	1.6
Completeness [%]	98.4	77.4	92.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		6.5	291	The protein complex was at 8 mg/ml in 10 mM Tris pH 7,5mm DTT. 2 microlitres drops were set up under paraffin oil. To 1.1 microlitres of precipitant (1.6 M sodium citrate pH 6.5, pH adjusted with HCl) 0.9 microlitres of protein was added, the final concentration of precipitant was therefore 0.88 M sodium citrate. For cryo crystals were passed through the 1.6 M sodium citrate pH 6.5 preciptant solution and then flash frozen in liquid nitrogen, microbatch under paraffin oil, temperature 291K