3BSN

Norwalk Virus polymerase bound to 5-nitrocytidine triphosphate and primer-template RNA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CLSI BEAMLINE 08ID-1
Synchrotron site	CLSI
Beamline	08ID-1
Temperature [K]	100
Detector technology	CCD
Collection date	2007-10-09
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97934
Spacegroup name	P 21 21 21
Unit cell lengths	74.310, 93.850, 96.030
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.710 - 1.800
R-factor	0.20251
Rwork	0.201
R-free	0.23585
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1sh0
RMSD bond length	0.006
RMSD bond angle	1.000
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	1.970
High resolution limit [Å]	1.800	1.800
Rmerge	0.079	0.614
Number of reflections	58959
<I/σ(I)>	13.7	2.3
Completeness [%]	93.6	78.3
Redundancy	4.8	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	298	16% (w/v) PEG 8000, 25% (w/v) glycerol, 100 mM Tris-HCl, 50 mM KCl, 4 mM MgCl2, 10 mM MnCl2, 14 mM 2-mercaptoethanol, 0.1% (w/v) CHAPS, 0.5 mM RNA duplex, 1 mM NTP, 0.03 mM polymerase, pH 7.0, VAPOR DIFFUSION, HANGING DROP, temperature 298K

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	PEG 8000
10	1	2	PEG 8000
11	1	2	glycerol
12	1	2	Tris-HCl
13	1	2	KCl
14	1	2	MgCl2
15	1	2	MnCl2
16	1	2	2-mercaptoethanol
17	1	2	CHAPS
18	1	2	NTP
2	1	1	glycerol
3	1	1	Tris-HCl
4	1	1	KCl
5	1	1	MgCl2
6	1	1	MnCl2
7	1	1	2-mercaptoethanol
8	1	1	CHAPS
9	1	1	NTP