1SI7

Structure of E. coli tRNA psi 13 pseudouridine synthase TruD

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X12C
Synchrotron site	NSLS
Beamline	X12C
Temperature [K]	93
Detector technology	CCD
Collection date	2003-11-15
Detector	CUSTOM-MADE
Wavelength(s)	0.97904
Spacegroup name	P 43
Unit cell lengths	63.479, 63.479, 112.235
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.000 * - 2.200
R-factor	0.212
Rwork	0.212
R-free	0.25300
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	unpublished TruD model SAD data on selenomethionine labeled protein
RMSD bond length	0.005
RMSD bond angle	22.400 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.280
High resolution limit [Å]	2.200	2.200
Rmerge	0.039	0.215
Total number of observations	88054 *
Number of reflections	22262
<I/σ(I)>	30.3	5.1
Completeness [%]	98.8	96.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8 *	298	MES, PEG 8000, ethylene glycol, pH 6.0, VAPOR DIFFUSION, HANGING DROP, temperature 298.0K
1	VAPOR DIFFUSION, HANGING DROP	8 *	298	MES, PEG 8000, ethylene glycol, pH 6.0, VAPOR DIFFUSION, HANGING DROP, temperature 298.0K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	5-10 (mg/ml)
2	1	drop	HEPES	20 (mM)	pH8.0
3	1	drop		250 (mM)
4	1	drop	EDTA	2 (mM)
5	1	drop	dithiothreitol	2 (mM)
6	1	drop	ethylene gylcol	25 (%(v/v))
7	1	reservoir	MES	0.1 (M)	pH6.0
8	1	reservoir	PEG8000	12-14 (%(w/v))
9	1	reservoir	ethylene glycol	25 (%(v/v))