1L9V

Non Structural protein encoded by gene segment 8 of rotavirus (NSP2), an NTPase, ssRNA binding and nucleic acid helix-destabilizing protein

Experimental procedure

Experimental method	MAD
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X8C
Synchrotron site	NSLS
Beamline	X8C
Temperature [K]	93
Detector technology	CCD
Collection date	2001-02-24
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9795
Spacegroup name	I 4 2 2
Unit cell lengths	108.432, 108.432, 152.877
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	40.000 * - 2.600
R-factor	0.24 *
Rwork	0.241
R-free	0.28840 *
Structure solution method	MAD
RMSD bond length	0.007
RMSD bond angle	1.250 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.690
High resolution limit [Å]	2.600	2.600
Rmerge	0.068	0.314 *
Number of reflections	75176
<I/σ(I)>	10.1
Completeness [%]	95.9 *	94.2
Redundancy	5.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.25	298	15% Peg 6000, 0.2 M Magnesium acetate, 0.1M Tris-HCl. Protein buffer 0.2M MgCl2, 0.5 mM EDTA , 0.5 mM DTT, 2mM Tris-HCl pH7.2. 0.2 M MgCl2 was necessary to make protein monodisperse. Protein:precipitant ratio 1:1. Plate like crystals (0.08 mm X 0.2 mm X 0.2 mm)Crystals grew in two to three weeks, pH 7.25, VAPOR DIFFUSION, HANGING DROP, temperature 298K
1	VAPOR DIFFUSION, HANGING DROP	7.25	298	15% Peg 6000, 0.2 M Magnesium acetate, 0.1M Tris-HCl. Protein buffer 0.2M MgCl2, 0.5 mM EDTA , 0.5 mM DTT, 2mM Tris-HCl pH7.2. 0.2 M MgCl2 was necessary to make protein monodisperse. Protein:precipitant ratio 1:1. Plate like crystals (0.08 mm X 0.2 mm X 0.2 mm)Crystals grew in two to three weeks, pH 7.25, VAPOR DIFFUSION, HANGING DROP, temperature 298K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	6-8 (mg/ml)
2	1	reservoir	peg6	12-15 (%)
3	1	reservoir	magnesium acetate	0.2 (M)
4	1	reservoir	Tris-HCl	100 (mM)	pH6.5-7.5