1QOQ
CRYSTAL STRUCTURE OF WILD-TYPE TRYPTOPHAN SYNTHASE COMPLEXED WITH INDOLE GLYCEROL PHOSPHATE
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X12C |
Synchrotron site | NSLS |
Beamline | X12C |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 1998-11-15 |
Detector | ADSC CCD |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 182.742, 60.055, 67.098 |
Unit cell angles | 90.00, 94.48, 90.00 |
Refinement procedure
Resolution | 20.000 - 1.800 |
R-factor | 0.171 * |
Rwork | 0.171 |
R-free | 0.21000 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1qop |
RMSD bond length | 0.017 |
RMSD bond angle | 0.033 |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | CNS |
Refinement software | REFMAC |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 31.200 | 1.900 |
High resolution limit [Å] | 1.800 | 1.800 |
Rmerge | 0.076 * | 0.209 * |
Total number of observations | 219018 * | |
Number of reflections | 63674 | |
<I/σ(I)> | 11.7 | 2.6 |
Completeness [%] | 94.5 | 70.8 |
Redundancy | 3.49 | 1.57 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7.8 | ENZYME SOLUTION: 10 MG/ML TRPS IN 50 MM BICINE PH 7.8, 1 MM EDTA, 5 MM DITHIOERYTHRITOL, 20 MUM PYRIDOXAL-5'-PHOSPHATE, RESERVOIR SOLUTION: 50 MM BICINE PH 7.8, 5 MM EDTA, 5 MM DITHIOERYTHRITOL, 0.1 MM PYRIDOXAL-5'-PHOSPHATE, 2 MM SPERMINE, 8-12 % PEG 8000, HANGING DROP GEOMETRY IN CONTRAST TO WHAT HAS BEEN IMPLICATED IN THE PUBLICATION ASSOCIATED WITH THIS PDB-ENTRY (WEYAND & SCHLICHTING, BIOCHEMISTRY 38: 16469-16480, (1999)), THE PH OF THE COMPLEX WAS NOT NEUTRAL BUT 5.0-5.2. THEREFORE, THE STRUCTURE WAS RE-DETERMINED AT PH 7.0 (PDB CODE 2RHG) AND ALSO AT PH 9.0 (PDB CODE 2RH9). THIS DATA SHOWS THAT CLOSURE OF LOOP ALPHA-L6 IS CAUSED BY THE LOW PH AND NOT BY BINDING OF IGP |
Crystallization Reagents in Literatures
ID | crystal ID | solution | reagent name | concentration (unit) | details |
1 | 1 | drop | protein | 10 (mg/ml) | |
10 | 1 | reservoir | spermine | 2 (mM) | |
11 | 1 | reservoir | 2 (mM) | ||
12 | 1 | reservoir | PEG8000 | 8-12 (%(w/v)) | |
2 | 1 | drop | Bicine | 50 (mM) | |
3 | 1 | drop | Na EDTA | 10 (mM) | |
4 | 1 | drop | di-thioerythritol | 1 (mM) | |
5 | 1 | drop | PLP | 0.020 (mM) | |
6 | 1 | reservoir | Bicine | 50 (mM) | |
7 | 1 | reservoir | di-thioerythritol | 5 (mM) | |
8 | 1 | reservoir | Na EDTA | 5 (mM) | |
9 | 1 | reservoir | PLP | 0.1 (mM) |