5LJF

Crystal structure of the endo-1,4-glucanase RBcel1 E135A with cellotriose

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 2
Synchrotron site	SOLEIL
Beamline	PROXIMA 2
Temperature [K]	100
Detector technology	CCD
Collection date	2015-06-11
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9801
Spacegroup name	P 21 21 21
Unit cell lengths	45.690, 99.310, 148.580
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.919 - 1.734
R-factor	0.177417744335
Rwork	0.176
R-free	0.21231
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4EE9 polyalanine
RMSD bond length	0.006
RMSD bond angle	0.920
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER (phenix.automr_1.10.1_2155)
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.090	1.800
High resolution limit [Å]	1.730	1.730
Rmerge	0.093	0.860
Number of reflections	70572
<I/σ(I)>	15.81	2.54
Completeness [%]	99.0	94
Redundancy	8.6	8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.4	292	Crystals were grown at 19 C using the hanging drop method by mixing 2 uL of protein solution (385 uM of protein in 20 mM NaPi pH 6.5 and cellotriose 5 mM) , with 2 uL of resevoir buffer (100 mM Tris HCl pH 7.4 with 17.5 pc w/v polyethylene glycol 600) containing seeds. Crystals were cryoprotected by equilibrating 2 hours the drop against 500 uL reservoir containing 0.1M Tris PEG 600 30pc pH 7.4.