2HOJ
Crystal structure of an E. coli thi-box riboswitch bound to thiamine pyrophosphate, manganese ions
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 5.0.2 |
| Synchrotron site | ALS |
| Beamline | 5.0.2 |
| Temperature [K] | 100 |
| Detector technology | AREA DETECTOR |
| Wavelength(s) | 1.0 |
| Spacegroup name | P 32 1 2 |
| Unit cell lengths | 64.700, 64.700, 101.900 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 30.000 - 2.500 |
| Rwork | 0.229 |
| R-free | 0.25700 |
| Structure solution method | MOLECULAR REPLACEMENT |
| RMSD bond length | 0.008 |
| RMSD bond angle | 1.370 |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | CNS |
| Refinement software | CNS |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 30.000 | 2.590 |
| High resolution limit [Å] | 2.500 | 2.500 |
| Rmerge | 0.062 | 0.548 |
| Number of reflections | 8398 | |
| Completeness [%] | 100.0 | 99.8 |
| Redundancy | 5.3 | 5.4 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 6 | 295 | grown: 26% PEG 2000, 10 mM MnCl2, 0.2 M NH4Cl, 50 mM cacodylate pH 6.0; 1.5 mM MgCl2, 0.5 mM TPP, 50 mM KCl, 0.25 mM spermine; cryo: above plus 15% Ficoll 400, VAPOR DIFFUSION, SITTING DROP, temperature 295K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | PEG 2000 | ||
| 10 | 1 | 2 | MnCl2 | ||
| 11 | 1 | 2 | NH4Cl | ||
| 12 | 1 | 2 | MgCl2 | ||
| 13 | 1 | 2 | KCl | ||
| 2 | 1 | 1 | MnCl2 | ||
| 3 | 1 | 1 | NH4Cl | ||
| 4 | 1 | 1 | MgCl2 | ||
| 5 | 1 | 1 | KCl | ||
| 6 | 1 | 1 | spermine | ||
| 7 | 1 | 1 | cacodylate | ||
| 8 | 1 | 1 | TPP | ||
| 9 | 1 | 2 | PEG 2000 |
