5D9O

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	CCD
Collection date	2014-10-22
Detector	RIGAKU SATURN A200
Wavelength(s)	1.5418
Spacegroup name	P 1 21 1
Unit cell lengths	49.180, 85.058, 74.978
Unit cell angles	90.00, 101.40, 90.00

Refinement procedure

Resolution	19.456 - 1.550
R-factor	0.1539
Rwork	0.152
R-free	0.18020
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3vdh
RMSD bond length	0.007
RMSD bond angle	1.118
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]	19.470	1.630
High resolution limit [Å]	1.550	1.550
Rmerge		0.535
Number of reflections	85184
<I/σ(I)>	14	2.9
Completeness [%]	97.1	94.1
Redundancy	4	4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		298	1.8 microL of protein solution at 28 mg/mL mixed with 1.8 microL of reservoir solution (0.1 M sodium cacodylate pH 6.3 to 7.1, 0.2 M calcium acetate, 25% PEG8K), then soaking crystals in reservoir solution supplemented with 10 mM cellotetraose for 2 h. Cryoprotectant = paratone-N oil.