1B97
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 | SYNCHROTRON |
| Source details | SRS BEAMLINE PX7.2 |
| Synchrotron site | SRS |
| Beamline | PX7.2 |
| Temperature [K] | 100 |
| Collection date | 1996-10-15 |
| Spacegroup name | P 1 |
| Unit cell lengths | 48.651, 49.224, 64.039 |
| Unit cell angles | 96.65, 108.91, 107.53 |
Refinement procedure
| Resolution | 15.000 - 1.900 |
| R-factor | 0.219 * |
| Rwork | 0.219 |
| R-free | 0.27900 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1rva |
| RMSD bond length | 0.013 |
| RMSD bond angle | 2.430 |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | AMoRE |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 15.000 | 1.990 |
| High resolution limit [Å] | 1.900 | 1.900 |
| Rmerge | 0.052 | 0.207 |
| Total number of observations | 113353 * | |
| Number of reflections | 38240 * | |
| <I/σ(I)> | 15.1 | 3.6 |
| Completeness [%] | 92.7 | 87.5 |
| Redundancy | 1.8 | 1.7 |
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 | DTT | ||
| 5 | 1 | 1 | NAN3 | ||
| 6 | 1 | 1 | SODIUM CACODYLATE | ||
| 7 | 1 | 1 | PEG 4000 | ||
| 8 | 1 | 2 | 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)) |
