4PSP

Crystal Structure of GH29 family alpha-L-fucosidase from Fusarium graminearum in the open form

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2012-03-08
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 1
Unit cell lengths	53.871, 75.887, 80.845
Unit cell angles	105.91, 107.23, 106.75

Refinement procedure

Resolution	41.235 - 1.561
R-factor	0.1364
Rwork	0.135
R-free	0.16910
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1hl8
RMSD bond length	0.009
RMSD bond angle	1.214
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	PHENIX (1.8.4_1496)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.590
High resolution limit [Å]	1.560	4.230	1.560
Rmerge	0.062	0.031	0.450
Number of reflections	148010
<I/σ(I)>	11.6
Completeness [%]	96.9	99.3	94.4
Redundancy	3.9	3.9	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	8	293.15	1:1 v/v mixture of 14-16 mg/ml alpha-L-fucosidase (stored in 25mM Tris pH 7.5 and partially deglycosylated by incubation 10:1:1 v/v ratio with EndoH and 500 mM sodium citrate pH 5.5 buffer from New England Biolabs for more than 24hrs before setting up the drop) with 30% PEG 2000mme, 0.1M Tris pH 8.0, crystals grow within two days, cryoprotected by Mitegen LV cryo-oil, batch method, temperature 293.15K, VAPOR DIFFUSION