3BSO

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	100
Detector technology	CCD
Collection date	2007-12-01
Detector	ADSC QUANTUM 315
Wavelength(s)	1.11587
Spacegroup name	P 21 21 21
Unit cell lengths	75.100, 93.473, 96.538
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	60.000 - 1.740
R-factor	0.20218
Rwork	0.201
R-free	0.23171
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.060
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	60.000	1.800
High resolution limit [Å]	1.740	1.740
Rmerge	0.045	0.436
Number of reflections	69074
<I/σ(I)>	29.2	2
Completeness [%]	98.1	90.5
Redundancy	3.8	3.2

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 CTP, 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	CTP
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	CTP