1BF3
P-HYDROXYBENZOATE HYDROXYLASE (PHBH) MUTANT WITH CYS 116 REPLACED BY SER (C116S) AND ARG 42 REPLACED BY LYS (R42K), IN COMPLEX WITH FAD AND 4-HYDROXYBENZOIC ACID
Experimental procedure
| Source type | ROTATING ANODE |
| Source details | MACSCIENCE M18X |
| Temperature [K] | 298 |
| Detector technology | AREA DETECTOR |
| Collection date | 1995-07 |
| Detector | SIEMENS |
| Spacegroup name | C 2 2 21 |
| Unit cell lengths | 71.350, 145.900, 88.800 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 8.000 - 2.200 |
| R-factor | 0.175 * |
| Rwork | 0.175 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1pbe |
| RMSD bond length | 0.009 |
| RMSD bond angle | 23.800 * |
| Data reduction software | XDS |
| Data scaling software | XSCALE |
| Phasing software | X-PLOR (3.8) |
| Refinement software | X-PLOR (3.8) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 8.000 | 2.000 |
| High resolution limit [Å] | 2.200 | 1.700 |
| Rmerge | 0.067 * | 0.045 |
| Total number of observations | 78553 * | |
| Number of reflections | 23174 | |
| <I/σ(I)> | 3 | |
| Completeness [%] | 96.8 | 82 |
| Redundancy | 2.5 | 2.5 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | Vapor diffusion, hanging drop * | 7 | 4 * | 39% AMMONIUMSULFATE, 100 MM SODIUM PHOSPHATE, 0.04 MM FAD, 0.15 MM EDTA, 30 MM SODIUM SULFITE, 1 MM P-HYDROXYBENZOATE, pH 7.0 |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | enzyme | 10-15 (mg/ml) | |
| 2 | 1 | drop | potassium phosphate | 100 (mM) | |
| 3 | 1 | reservoir | ammonium sulfate | 39 (%sat) | |
| 4 | 1 | reservoir | FAD | 0.04 (mM) | |
| 5 | 1 | reservoir | EDTA | 0.15 (mM) | |
| 6 | 1 | reservoir | sodium sulfite | 30 (mM) | |
| 7 | 1 | reservoir | 4-hydroxybenzoate | 1 (mM) | |
| 8 | 1 | reservoir | potassium phosphate | 100 (mM) |
