2OC8

Structure of Hepatitis C Viral NS3 protease domain complexed with NS4A peptide and ketoamide SCH503034

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	95
Detector technology	CCD
Detector	ADSC QUANTUM 210
Wavelength(s)	1.00
Spacegroup name	H 3 2
Unit cell lengths	223.871, 223.871, 74.962
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	8.000 - 2.660
Rwork	0.192
R-free	0.27800
Structure solution method	MIR
Starting model (for MR)	2o8m
RMSD bond length	0.008
RMSD bond angle	1.854
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
High resolution limit [Å]	2.650	2.650
Number of reflections	20317
<I/σ(I)>	31.6	3.5
Completeness [%]	97.1	70
Redundancy	5.3	5.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6	277	The protein (NS3 complexed with KK-NS4a(21-39)-KK peptide) was at 12-15 mg/ml in 15 mM MES, pH 6.5, 1 M NaCl, 20 mM b-mercaptoethanol. Hanging Drops were formed by mixing 4:l protein solution with 4:l {0.75-1.0 M NaCl, 0.1M Na/K phosphate, 0.1 M Mes, pH 5.8-6.1, 20 mM 2-mercaptoethanol} The drop was equilibrated the drops over 1 ml {(1.25-1.50 M) NaCl - 0.1M Na/K phosphate 0.1 M Mes, pH 5.6-5.8, 20 mM 2-mercaptoethanol} , VAPOR DIFFUSION, HANGING DROP, temperature 277K