1DY9
Inhibition of the Hepatitis C Virus NS3/4A Protease. The Crystal Structures of Two Protease-Inhibitor Complexes (inhibitor I)
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE ID14-3 |
| Synchrotron site | ESRF |
| Beamline | ID14-3 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 1999-02-15 |
| Detector | MARRESEARCH |
| Spacegroup name | P 61 |
| Unit cell lengths | 94.350, 94.350, 82.370 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 20.000 - 2.100 |
| R-factor | 0.207 |
| Rwork | 0.213 |
| R-free | 0.27500 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1dy7 |
| RMSD bond length | 0.010 |
| RMSD bond angle | 0.030 |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | AMoRE |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 20.000 | 2.140 |
| High resolution limit [Å] | 2.100 | 2.100 |
| Rmerge | 0.035 * | 0.150 * |
| Total number of observations | 66924 * | |
| Number of reflections | 23960 | |
| <I/σ(I)> | 32 | 3.2 |
| Completeness [%] | 98.2 | 98.4 |
| Redundancy | 2.8 | 2.1 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | Vapor diffusion, hanging drop * | 5.1 | THE NS3 PROTEIN (1MG/ML) WAS INCUBATE 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. THE TERNARY COMPLEX WITH INHIBITOR WAS PREPARED BY ADDING 5.0 MM OF INIBITOR TO CRYSTALS THAT WERE STABILISED IN 4.5 M NACL, 10 MM 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) |
