2HOM
Crystal structure of an E. coli thi-box riboswitch bound to thiamine monophosphate
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 8.2.2 |
| Synchrotron site | ALS |
| Beamline | 8.2.2 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2005-07-10 |
| Wavelength(s) | 0.95 |
| Spacegroup name | P 32 1 2 |
| Unit cell lengths | 65.700, 65.700, 101.700 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 30.000 - 2.890 |
| Rwork | 0.203 |
| R-free | 0.26600 |
| Structure solution method | MOLECULAR REPLACEMENT |
| RMSD bond length | 0.006 |
| RMSD bond angle | 1.280 |
| 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 | 2.950 |
| High resolution limit [Å] | 2.890 | 2.890 |
| Rmerge | 0.124 | 0.390 |
| Number of reflections | 5263 | |
| <I/σ(I)> | 16.4 | 2.1 |
| Completeness [%] | 92.6 | 52.6 |
| Redundancy | 5.4 | 2.6 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 7 | 295 | growth: 29% PEG 2000, 15 mM CaCl2, 0.2 M NH4Cl, 50 mM succinate pH 7; 0.25 mM TMP, 0.25 mM spermine, 1.5 mM MgCl2, 50 mM KCl; cryo: above with 15% w/v sucrose; 500 x 70 x 70 um crystal, pH 7.0, 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 | PEG 2000 | ||
| 11 | 1 | 2 | CaCl2 | ||
| 12 | 1 | 2 | NH4Cl | ||
| 13 | 1 | 2 | MgCl2 | ||
| 14 | 1 | 2 | KCl | ||
| 15 | 1 | 2 | succinate | ||
| 2 | 1 | 1 | CaCl2 | ||
| 3 | 1 | 1 | NH4Cl | ||
| 4 | 1 | 1 | MgCl2 | ||
| 5 | 1 | 1 | KCl | ||
| 6 | 1 | 1 | succinate | ||
| 7 | 1 | 1 | spermine | ||
| 8 | 1 | 1 | cacodylate | ||
| 9 | 1 | 1 | TMP |
