1B96
ANALYSIS OF A MUTATIONAL HOT-SPOT IN THE ECORV RESTRICTION ENDONUCLEASE: A CATALYTIC ROLE FOR A MAIN CHAIN CARBONYL GROUP
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | ROTATING ANODE |
| Source details | SIEMENS |
| Temperature [K] | 100 |
| Detector technology | IMAGE PLATE |
| Collection date | 1997-01-15 |
| Detector | MACSCIENCE |
| Spacegroup name | P 1 |
| Unit cell lengths | 48.350, 48.780, 63.640 |
| Unit cell angles | 96.61, 108.94, 107.44 |
Refinement procedure
| Resolution | 12.500 - 2.300 |
| R-factor | 0.189 * |
| Rwork | 0.189 |
| R-free | 0.28200 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1rva |
| RMSD bond length | 0.011 |
| RMSD bond angle | 2.426 |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | AMoRE |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 12.500 | 2.380 |
| High resolution limit [Å] | 2.300 | 2.300 |
| Rmerge | 0.099 | 0.303 |
| Total number of observations | 148523 * | |
| Number of reflections | 21703 | |
| <I/σ(I)> | 7.1 | 1.6 |
| Completeness [%] | 96.5 | 94.8 |
| Redundancy | 1.56 | 1.61 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 7.5 * | 18 * | pH 7.0, VAPOR DIFFUSION, SITTING DROP |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | PHOSPHATE BUFFER | ||
| 2 | 1 | 1 | NACL | ||
| 3 | 1 | 1 | EDTA | ||
| 4 | 1 | 1 | NAN3 | ||
| 5 | 1 | 1 | DTT | ||
| 6 | 1 | 1 | SODIUM CACODYLATE | ||
| 7 | 1 | 1 | PEG 4000 |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | protein | 15 (mg/ml) | 0.260mM dimer |
| 2 | 1 | drop | phosphate | 10 (mM) | pH7.5 |
| 3 | 1 | drop | 250 (mM) | ||
| 4 | 1 | drop | EDTA | 1 (mM) | |
| 5 | 1 | drop | dithiothreitol | 0.1 (mM) | |
| 6 | 1 | drop | sodium azide | 0.1 (%(w/v)) | |
| 7 | 1 | drop | DNA | 3 (mg/ml) | 0.460mM |
| 8 | 1 | drop | cacodylic | 10 (mM) | pH6.0 |
| 9 | 1 | drop | PEG4000 | 0.8-1.4 (%(w/v)) |
