6WQY

GH5-4 broad specificity endoglucanase from Bacteroides salanitronis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-10-20
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.7749
Spacegroup name	P 21 21 21
Unit cell lengths	40.480, 92.310, 95.150
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.240 - 1.050
R-factor	0.1389
Rwork	0.139
R-free	0.14530
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4yhe
RMSD bond length	0.005
RMSD bond angle	0.882
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]	29.240	1.088
High resolution limit [Å]	1.050	1.050
Rmerge	0.051	0.832
Rmeas	0.053	0.911
Rpim	0.015	0.358
Number of reflections	165260	15327
<I/σ(I)>	22.62	1.68
Completeness [%]	99.2	93
Redundancy	12.1	6.1
CC(1/2)	1.000	0.621

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	293	200 nL of a solution containing 59 mg/mL protein, 50 nM NaCl, 5 mM MOPS buffer pH 7 was mixed with 130 nL reservoir solution containing 16% PEG3350, 0.2 M MgCl2, and 0.1 M sodium acetate buffer at pH 5. Vapor diffusion in MRC plates. Crystals appeared within a few days, and were cryo-protected using a reservoir solution supplemented to 35% PEG3350.