4PSR

Crystal Structure of alpha-L-fucosidase from Fusarium graminearum in the open form in complex with L-fucose

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	2012-01-01
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 1
Unit cell lengths	54.153, 75.989, 80.970
Unit cell angles	105.69, 107.32, 106.93

Refinement procedure

Resolution	43.280 - 1.380
R-factor	0.1397
Rwork	0.138
R-free	0.16610
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1hl8
RMSD bond length	0.011
RMSD bond angle	1.381
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	1.400
High resolution limit [Å]	1.380	3.750	1.380
Rmerge	0.062	0.029	0.623
Number of reflections	214983
<I/σ(I)>	9.8
Completeness [%]	96.1	99.2	93.2
Redundancy	3.9	3.9	3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	batch	8	293.15	11:1 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, soaked with 50 mM fucose (final concentration) in crystallization solution for around 5 minutes, cryoprotected by Mitegen LV cryo-oil, batch method, temperature 293.15K), flash frozen in liquid nitrogen before data collection.