3IHP

Covalent Ubiquitin-Usp5 Complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	CCD
Collection date	2007-06-11
Detector	MARMOSAIC 300 mm CCD
Spacegroup name	P 21 21 21
Unit cell lengths	68.295, 188.845, 207.860
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	34.900 - 2.800
R-factor	0.22688
Rwork	0.224
R-free	0.27588
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	PDB ENTRIES 2IBI 2g43 2dak 2dag
RMSD bond length	0.011
RMSD bond angle	1.308
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.000	2.900
High resolution limit [Å]	2.800	2.800
Number of reflections	66229
<I/σ(I)>	17.1	2
Completeness [%]	98.0	94.6
Redundancy	5.4	5.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		6.5	298	Crystals of the covalent ubiquitin complex of Usp5 were grown at 298 K using the hanging drop method by mixing equal volumes of protein solution (25 mg/ml) and Crystallization Buffer (1.45 M ammonium sulfate, 0.1 M bis-Tris, pH 6.5, 0.2 M sodium acetate, 5% ethyleneglycol and 1 mM dithiothreitol). The crystals were cryoprotected by immersion in Paratone N in paraffin oil 30% (v/v) and placed in liquid nitrogen., VAPOR DIFFUSION, HANGING DROP, temperature 298.0K