1SH2

Crystal Structure of Norwalk Virus Polymerase (Metal-free, Centered Orthorhombic)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	110
Detector technology	IMAGE PLATE
Collection date	2003-10-01
Detector	MARRESEARCH
Wavelength(s)	1.5418
Spacegroup name	C 2 2 21
Unit cell lengths	114.270, 115.240, 91.900
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	40.000 * - 2.300
R-factor	0.23054
Rwork	0.227
R-free	0.29300 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1khv
RMSD bond length	0.007 *
RMSD bond angle	1.080 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	BEAST
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.370
High resolution limit [Å]	2.300	2.300
Rmerge	0.075	0.402
Total number of observations	215719 *
Number of reflections	27003	2054 *
<I/σ(I)>	19.8	2.3
Completeness [%]	98.8	91.3
Redundancy	8	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	295	PEG 8000, ammonium sulfate, Tris-Cl, glycerol, CHAPS, 2-mercaptoethanol, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 295K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	polymerase	7 (mg/ml)
2	1	reservoir	PEG8000	24 (%(w/v))
3	1	reservoir	ammonium sulfate	100-200 (mM)
4	1	reservoir	Tris-Cl	50 (mM)	pH7.5
5	1	reservoir	glycerol	15 (%(w/v))
6	1	reservoir	CHAPS	0.2 (%(w/v))
7	1	reservoir	2-mercaptoethanol	14 (mM)