7TDA

Crystal structure of the E. coli thiM riboswitch in complex with thiamine pyrophosphate, 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	65.410, 65.410, 101.504
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	29.050 - 2.250
R-factor	0.2112
Rwork	0.209
R-free	0.23130
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2hoj
RMSD bond length	0.004
RMSD bond angle	0.951
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX ((1.14_3260))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.290
High resolution limit [Å]	2.250	2.250
Rmerge	0.064	1.297
Rpim	0.019	0.437
Number of reflections	12071	589
<I/σ(I)>	82.7	1
Completeness [%]	100.0	100
Redundancy	12.4	9.6
CC(1/2)	0.994	0.700

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		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 0.5 mM of TPP. the reservoir solution was 50 mM Bis-Tris HCl, pH 6.5, 0.5 M NH4Cl, 10 mm MnCl2, and 30% (v/v) PEG2000