2OML

crystal structure of E. coli pseudouridine synthase RluE

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	100
Detector technology	CCD
Collection date	2003-06-02
Detector	ADSC QUANTUM 4
Spacegroup name	P 21 21 21
Unit cell lengths	32.310, 56.780, 91.310
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	28.080 - 1.200
R-factor	0.152
Rwork	0.149
R-free	0.17900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1KSV with N-terminal domain removed
RMSD bond length	0.014
RMSD bond angle	1.672
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	1.240
High resolution limit [Å]	1.200	1.200
Rmerge	0.063	0.713
Number of reflections	52181
<I/σ(I)>	13.7	2.1
Completeness [%]	98.1	94
Redundancy	4.6	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	294	4 mg/ml protein in 10 mM Tris, pH 7.5, 2mM EDTA and 2mM DTT equilibrated against 22% (w/v) MME PEG 2000, 200 mM ammonium sulfate, 100 mM sodium acetate, crystals were improved by microseeding a solution of 18-22% MME PEG 2000, 200 mM ammonium sulfate, 100 mM sodium acetate and 2 mg/ml protein at pH 5.0, VAPOR DIFFUSION, HANGING DROP, temperature 294K