2HOL
Crystal structure of an E. coli thi-box riboswitch bound to thiamine pyrophosphate, barium ions
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ALS BEAMLINE 5.0.1 |
Synchrotron site | ALS |
Beamline | 5.0.1 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2005-10-06 |
Spacegroup name | P 32 1 2 |
Unit cell lengths | 65.500, 65.500, 101.700 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 30.000 - 2.900 |
R-factor | 0.21 |
Rwork | 0.210 |
R-free | 0.24400 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.020 |
RMSD bond angle | 2.310 |
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 | 3.000 |
High resolution limit [Å] | 2.900 | 2.900 |
Rmerge | 0.098 | 0.457 |
Number of reflections | 5517 | |
<I/σ(I)> | 25.5 | 3.8 |
Completeness [%] | 100.0 | 99.9 |
Redundancy | 13.3 | 11 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | 6 | 295 | growth: 23% PEG 3350, 0.2 M NH4Cl, 10 mM CaCl2, 50 mM Cacodylate; 0.25 mM TPP, 0.25 mM spermine, 50 mM KCl, 1.5 mM MgCl2; soaked in 60 mM BaCl2 for 7 days; cryo above with 20% w/v sucrose and 26% PEG 2000 rather than 3350, pH 6.0, VAPOR DIFFUSION, HANGING DROP, temperature 295K, pH 6.00 |
Crystallization Reagents
ID | crystal ID | solution ID | reagent name | concentration | details |
1 | 1 | 1 | PEG 3350 | ||
10 | 1 | 2 | PEG 3350 | ||
11 | 1 | 2 | CaCl2 | ||
12 | 1 | 2 | NH4Cl | ||
13 | 1 | 2 | MgCl2 | ||
14 | 1 | 2 | KCl | ||
15 | 1 | 2 | sucrose | ||
16 | 1 | 3 | BaCl2 | ||
2 | 1 | 1 | CaCl2 | ||
3 | 1 | 1 | NH4Cl | ||
4 | 1 | 1 | MgCl2 | ||
5 | 1 | 1 | KCl | ||
6 | 1 | 1 | sucrose | ||
7 | 1 | 1 | spermine | ||
8 | 1 | 1 | cacodylate | ||
9 | 1 | 1 | TPP |