4WTJ

CRYSTAL STRUCTURE OF HCV NS5B GENOTYPE 2A JFH-1 ISOLATE WITH S15G E86Q E87Q C223H V321I MUTATIONS IN COMPLEX WITH RNA TEMPLATE 5'-AUCC, RNA PRIMER 5'-PGG, MN2+, AND ADP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	100
Detector technology	CCD
Collection date	2013-02-17
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.12709
Spacegroup name	P 65
Unit cell lengths	139.870, 139.870, 92.700
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.730 - 2.200
R-factor	0.178
Rwork	0.176
R-free	0.21100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4obc
RMSD bond length	0.011
RMSD bond angle	1.489
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.7.0032)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.260
High resolution limit [Å]	2.200	2.200
Rmerge	0.074	0.643
Rmeas	0.080	0.019
Total number of observations	350400
Number of reflections	52305	539
<I/σ(I)>	23.15	3.6
Completeness [%]	99.8	100
Redundancy	6.7	6.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.6	289	NS5B AT 4.7 MG/ML IN 5 MM TRIS PH 7.5, 200 MM NH4OAC, 1 MM EDTA, 1 MM DTT AGAINST 25% PEG 550 MME, 50 MM MGCL2, 0.1 M HEPES PH 7.5 FOR CRYSTAL GROWTH SOAKED INTO 28% PEG 550 MME, 0.2 M AMMONIUM ACETATE, 0.05 M BISTRIS PROPANE PH 6.0, 0.05 M TRIS PH 7.2, 6 MM MNCL2, 10 MM CDP, 2 MM 5'-AGCC, 2 MM 5'-PGG, WITH 8% GLYCEROL AS CRYO-PROTECTANT, CRYSTAL TRACKING ID 251148A2, UNIQUE PUCK ID HDW8-4