1DXP

Inhibition of the Hepatitis C Virus NS3/4A Protease. The Crystal Structures of Two Protease-Inhibitor Complexes (apo structure)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE X11
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	X11
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1999-04-15
Detector	MARRESEARCH
Spacegroup name	P 61
Unit cell lengths	92.980, 92.980, 81.810
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	20.000 - 2.400
R-factor	0.196
Rwork	0.198
R-free	0.29800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1jxp
RMSD bond length	0.010
RMSD bond angle	0.037
Data reduction software	DENZO
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.530
High resolution limit [Å]	2.400	2.400
Rmerge	0.071 *	0.355 *
Total number of observations	126238 *
Number of reflections	15786
<I/σ(I)>	9	2.2
Completeness [%]	100.0	99.9
Redundancy	8	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	5.1		THE NS3 PROTEIN (1MG/ML) WAS INCUBATED AT 4C WITH THE NS4A COFACTOR PEPTIDE, CONTAINING A SOLUBIZING LYSINE TAG AT ITS N- AND C-TERMINI(KGSVVIVGRIILSGRK), AT A MOLAR RATIO OF 1:2 AND CONCENTRATED TO 290 MICROMOLAR. NS3J/4A CRYSTALS, WITH A MAXIMUM SIZE OF 0.6 X 0.3 X 0.2 MM**3, WERE OBTAINED BY BOTH HANGING- AND SITTING-DROP VAPOUR DIFFUSION METHODS AFTER TWO WEEKS AT ROOM TEMPERATURE, WITH 3.4 M NACL, 10MM DTT, 0.1 M CITRATE BUFFER PH 5.1.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir		3.4 (M)
2	1	reservoir	cyclohexyl-pentyl-beta-D-maltoside	4.8 (mM)
3	1	reservoir	dithiothreitol	5 (mM)
4	1	reservoir		0.02 (%)
5	1	reservoir	citrate	0.1 (M)
6	1	drop		4.5 (M)
7	1	drop	dithiothreitol	10 (mM)
8	1	drop	citrate	0.1 (M)