4XT0

Crystal Structure of Beta-etherase LigF from Sphingobium sp. strain SYK-6

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2014-02-05
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97857
Spacegroup name	P 63 2 2
Unit cell lengths	123.708, 123.708, 66.416
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	40.493 - 2.070
R-factor	0.161
Rwork	0.158
R-free	0.21430
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.022
Data scaling software	SCALEPACK
Phasing software	PHASER (2.5.5)
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	40.493	50.000	2.110
High resolution limit [Å]	2.070	5.620	2.070
Rmerge	0.076	0.038	0.637
Rmeas	0.079	0.039	0.663
Rpim	0.019	0.010	0.181
Total number of observations	326246
Number of reflections	18884
<I/σ(I)>	11.9
Completeness [%]	99.8	98.8	98.3
Redundancy	17.3	15.5	12.9
CC(1/2)		0.998	0.922

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		298	1 uL protein solution (10mM HEPES pH 7.5, 50mM NaCl, 0.5mM TCEP, 1mM glutathione) mixed with 0.8 uL of the well solution (25% MEPEG 2000, 0.2M TMAO, 0.1M BTP pH 9) and 0.2 uL of a suspension of LigF micro-crystals (0.2M Magnesium formate, 30% PEG 3350, 1mM glutathione) Cryoprotected with 30% MEPEG 2000, 0.2M TMAO, 0.1M Tris pH 8.5, 1mM glutathione