6MQ4

GH5-4 broad specificity endoglucanase from Hungateiclostridium cellulolyticum

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2015-08-23
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97857
Spacegroup name	P 31
Unit cell lengths	70.850, 70.850, 57.610
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	30.176 - 1.400
R-factor	0.1213
Rwork	0.121
R-free	0.13770
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2wab
RMSD bond length	0.010
RMSD bond angle	1.174
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((1.14_3211: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.176	1.450
High resolution limit [Å]	1.400	1.400
Rmerge	0.053	0.423
Rmeas	0.058	0.458
Rpim	0.022	0.175
Number of reflections	63716	6406
<I/σ(I)>	22.1	4.3
Completeness [%]	100.0	100
Redundancy	7	6.9
CC(1/2)	0.999	0.921

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.2	293	A solution of protein at 1mM was incubated with 5mM cellohexose for 4 hours at room temperature. The mixture was screened using a TTP Labtech Mosquito and MRC SD-2 plates 200 nL protein 200 nL reservoir, 50 microliter total reservoir. Crystals formed in condition B6, 40% ethanol, 10% PEG1000, 0.1M phosphate citrate buffer pH 4.2. Crystals were exposed to vapor from 45% ethanol for 30 seconds prior to plunge cooling in liquid nitrogen.