7TD7

Crystal structure of an E. coli thiM riboswitch bound to thiamine, manganese ions

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-2
Synchrotron site	NSLS-II
Beamline	17-ID-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-02-01
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9793
Spacegroup name	P 32 1 2
Unit cell lengths	61.110, 61.110, 103.416
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	29.300 - 2.950
R-factor	0.2353
Rwork	0.230
R-free	0.28560
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2hoj
RMSD bond length	0.005
RMSD bond angle	1.076
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER (1.14_3260)
Refinement software	PHENIX (1.14_3260)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	3.060
High resolution limit [Å]	2.950	2.950
Rmerge	0.111	0.555
Rpim	0.043	0.237
Number of reflections	4766	445
<I/σ(I)>	47.6	1.6
Completeness [%]	98.8	93.3
Redundancy	8.4	5.3
CC(1/2)	0.985	0.933

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	291	RNA (0.15 mM) was incubated in a buffer containing 5 mM Tris-HCl, pH 8.0, 3 mM MgCl2, 10 mM NaCl, 0.1 M KCl, and 0.5 mM spermine with 1.0 mM of thiamine. the reservoir solution was 50 mM Bis-Tris HCl, pH 6.5, 0.5 M NH4Cl, 10 mm MnCl2, and 30% (v/v) PEG2000