117E
THE R78K AND D117E ACTIVE SITE VARIANTS OF SACCHAROMYCES CEREVISIAE SOLUBLE INORGANIC PYROPHOSPHATASE: STRUCTURAL STUDIES AND MECHANISTIC IMPLICATIONS
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | EMBL/DESY, HAMBURG BEAMLINE BW7A |
| Synchrotron site | EMBL/DESY, HAMBURG |
| Beamline | BW7A |
| Temperature [K] | 100 |
| Detector technology | IMAGE PLATE |
| Collection date | 1996-09 |
| Detector | MARRESEARCH |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 58.400, 102.800, 116.700 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 8.000 - 2.150 |
| R-factor | 0.182 |
| Rwork | 0.182 |
| R-free | 0.24000 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1wgj |
| RMSD bond length | 0.007 |
| RMSD bond angle | 26.200 * |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | X-PLOR |
| Refinement software | X-PLOR (3.851) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 20.000 | 2.190 |
| High resolution limit [Å] | 2.150 | 2.150 |
| Rmerge | 0.086 | |
| Total number of observations | 228918 * | |
| Number of reflections | 36385 | |
| <I/σ(I)> | 5.7 | |
| Completeness [%] | 88.9 | 86 |
| Redundancy | 6.3 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | Vapor diffusion, sitting drop * | 6 | 4 * | 17-19% MPD, 25 MM MES, PH 6.0, 1 MM MNCL2, 0.5 MM NA2HPO4, 10 MG/ML PROTEIN |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | protein | 10 (mg/ml) | |
| 2 | 1 | drop | MES | 25 (mM) | |
| 3 | 1 | drop | 1 (mM) | ||
| 4 | 1 | drop | sodium hydrogen phosphate | 0.5 (mM) | |
| 5 | 1 | drop | MPD | 17 (%(v/v)) | |
| 6 | 1 | reservoir | MPD | 19 (%(v/v)) |
