3KNX

HCV NS3 protease domain with P1-P3 macrocyclic ketoamide inhibitor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2004-04-01
Detector	RIGAKU RAXIS IV
Wavelength(s)	1.54
Spacegroup name	H 3 2
Unit cell lengths	223.587, 223.587, 75.282
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	8.000 - 2.650
Rwork	0.169
R-free	0.26300
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	HCV NS3(Ser139A)-NS4a peptide
RMSD bond length	0.008
RMSD bond angle	1.948
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (98.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.660
High resolution limit [Å]	2.600	2.600
Rmerge	0.089	0.395
Number of reflections	18845
<I/σ(I)>	12.8	1.5
Completeness [%]	84.8	13.5
Redundancy	4.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6	285	Crystallization was performed by the vapor diffusion method using hanging drops (4 L protein solution mixed with 4 L (0.75 - 1.00) M NaCl - 0.1 M MES - 0.1 M Na/K PO4, pH 5.6 - 6.2) suspended over 1 mL reservoir solutions of (1.25 - 1.50) M NaCl - 0.1 M MES - 0.1 M Na/K PO4 - 5 mM -mercaptoethanol, pH 5.6-6.2. The trays were set at 4oC for 5-7 days to control nucleation, followed by incubation for 3 weeks at 12oC to maximize crystal growth. , VAPOR DIFFUSION, HANGING DROP, temperature 277, then 285K