5D9M

Crystal structure of PbGH5A, a glycoside hydrolase family 5 enzyme from Prevotella bryantii B14, E280A mutant in complex with the xyloglucan tetradecasaccharide XXXGXXXG

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	CCD
Collection date	2014-11-19
Detector	RIGAKU SATURN A200
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	77.431, 81.677, 108.492
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	36.463 - 1.900
R-factor	0.1738
Rwork	0.173
R-free	0.20390
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3vdh
RMSD bond length	0.005
RMSD bond angle	0.915
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHENIX
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.960
High resolution limit [Å]	1.900	1.900
Rmerge		0.501
Number of reflections	54672
<I/σ(I)>	23.51	3.88
Completeness [%]	99.2	98.3
Redundancy	7.5	7.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	298	protein solution at 28 mg/mL mixed with 2.4 mM ligand incubated at 37 degrees C for 3 h, mixed with reservoir solution (0.2 M magnesium acetate, 20% (w/v) PEG3350). Cryoprotectant = paratone-N oil.