1B4E
X-ray structure of 5-aminolevulinic acid dehydratase complexed with the inhibitor levulinic acid
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | SRS BEAMLINE PX9.6 |
| Synchrotron site | SRS |
| Beamline | PX9.6 |
| Temperature [K] | 100 |
| Detector technology | IMAGE PLATE |
| Collection date | 1994-03-15 |
| Detector | MAR scanner 300 mm plate |
| Spacegroup name | I 4 2 2 |
| Unit cell lengths | 126.700, 126.700, 141.600 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 7.700 * - 2.000 |
| Rwork | 0.188 |
| R-free | 0.25500 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | YEAST ALAD |
| RMSD bond length | 0.016 |
| RMSD bond angle | 0.032 |
| Data reduction software | MOSFLM |
| Data scaling software | CCP4 ((AGROVATA) |
| Phasing software | CCP4 ((ALMN) |
| Refinement software | RESTRAIN |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 7.700 * | 2.100 |
| High resolution limit [Å] | 2.000 | 2.000 |
| Rmerge | 0.063 | 0.297 |
| Number of reflections | 33275 | |
| <I/σ(I)> | 8.8 | 2.1 |
| Completeness [%] | 85.8 | 70.8 |
| Redundancy | 2.5 | 1.6 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 8.2 | 7MG/ML ENZYME AND 2 - 5 % SATURATED AMMONIUM SULPHATE AS PRECIPITANT. 0.2 M TRIS-HCL USED TO BUFFER THE PH BETWEEN 8.1 AND 8.4. 15MM LEVULINIC ACID, 40UM ZINC SULPHATE AND 4MM BETA- MERCAPTOETHANOL WERE PRESENT., pH 8.2, VAPOR DIFFUSION, HANGING DROP |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | enzyme | 7 (mg/ml) | |
| 2 | 1 | drop | Tris-HCl | 0.2 (M) | |
| 3 | 1 | drop | levulinic acid | 15 (mM) | |
| 4 | 1 | drop | zinc sulfate | 0.04 (M) | |
| 5 | 1 | drop | mercaptoethanol | 4 (mM) | |
| 6 | 1 | reservoir | ammonium sulfate | 2-5 (%sat) |
