4LX4

Crystal Structure Determination of Pseudomonas stutzeri endoglucanase Cel5A using a Twinned Data Set

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM30A
Synchrotron site	ESRF
Beamline	BM30A
Temperature [K]	100
Detector technology	CCD
Collection date	2012-05-18
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.979639
Spacegroup name	P 1 21 1
Unit cell lengths	70.390, 82.900, 104.740
Unit cell angles	90.00, 92.08, 90.00

Refinement procedure

Resolution	48.279 - 1.556
R-factor	0.2236
Rwork	0.220
R-free	0.23710
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4ee9
RMSD bond length	0.007
RMSD bond angle	1.177
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER ((phenix.automr: 1.8.2_1309))
Refinement software	PHENIX ((phenix.refine: 1.8.2_1309))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.279	1.600
High resolution limit [Å]	1.556	1.560
Rmerge	0.076	0.339
Number of reflections	157658
<I/σ(I)>	17.9	5.9
Completeness [%]	91.6	97.7
Redundancy	7.1	7.07

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	292	Ps_Cel5A (27 microM) in 50 mM sodium phosphate pH 7.0, was mixed 1:1 with well buffer (100 mM tris pH 5.9 with 22.5 % v/v polyethylene glycol 600) using the hanging drop method with 500 uL well buffer in the well of the crystallization tray. Crystals obtained by this method were soaked 1 hour in 100 mM Tris pH 5.9 with 30% v/v polyethylene glycol 600., VAPOR DIFFUSION, HANGING DROP, temperature 292K