1K8W

Crystal structure of the E. coli pseudouridine synthase TruB bound to a T stem-loop RNA

Experimental procedure

Experimental method	MAD
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	CCD
Collection date	2001-06-07
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9611, 0.9792, 0.9794
Spacegroup name	C 1 2 1
Unit cell lengths	145.054, 40.361, 77.987
Unit cell angles	90.00, 110.60, 90.00

Refinement procedure

Resolution	28.270 - 1.850
R-factor	0.184
Rwork	0.184
R-free	0.21100
Structure solution method	MAD
RMSD bond length	0.006
RMSD bond angle	1.300
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	1.920
High resolution limit [Å]	1.850	1.850
Rmerge	0.062 *	0.326 *
Total number of observations	107243 *
Number of reflections	35366
Completeness [%]	96.8	94.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	22 *	ammonium sulfate, magnesium chloride, peg400, hepes, tcep, pH 7.5, VAPOR DIFFUSION, SITTING DROP, temperature 295K
1	VAPOR DIFFUSION, SITTING DROP	7.5	22 *	ammonium sulfate, magnesium chloride, peg400, hepes, tcep, pH 7.5, VAPOR DIFFUSION, SITTING DROP, temperature 295K

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	(NH4)2SO4
2	1	1	MgCl2
3	1	1	PEG 400
4	1	1	HEPES
5	1	1	TCEP

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	0.21 (mM)
2	1	reservoir	ammonium sulfate	1.5 (M)
3	1	reservoir		10 (mM)
4	1	reservoir	PEG400	2 (%)
5	1	reservoir	HEPES-KOH	0.1 (M)	pH7.5
6	1	reservoir	TCEP	1 (mM)