4NI3

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2013-04-20
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	P 1
Unit cell lengths	53.511, 75.512, 80.367
Unit cell angles	105.61, 107.19, 106.71

Refinement procedure

Resolution	37.931 - 1.399
R-factor	0.1449
Rwork	0.145
R-free	0.16670
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1HL8 (peptide only)
RMSD bond length	0.010
RMSD bond angle	1.248
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.8.4_1496)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	37.931	1.420
High resolution limit [Å]	1.399	1.399
Number of reflections	198748
<I/σ(I)>		2.3
Completeness [%]	94.9	92
Redundancy		3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	batch method	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 40% PEG 2000mme, 0.1M Tris pH 8.0, crystals grow within two days, cryoprotected by Mitegen LV cryo-oil, batch method, temperature 293.15K