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