6OHE

Alpha-L-fucosidase AlfC D200A in complex with Fuca(1,6)GlcNAc

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-2
Synchrotron site	NSLS-II
Beamline	17-ID-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-02-15
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.979330
Spacegroup name	C 2 2 21
Unit cell lengths	89.022, 139.251, 263.375
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.760 - 3.140
R-factor	0.2795
Rwork	0.279
R-free	0.29860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6o18
RMSD bond length	0.007
RMSD bond angle	1.259
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	MOLREP
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	29.760	29.760	3.330
High resolution limit [Å]	3.140	9.420	3.140
Rmerge	0.189	0.070	1.568
Rmeas	0.211	0.079	1.755
Rpim	0.092	0.035	0.775
Total number of observations	143330
Number of reflections	28950	1116	4624
<I/σ(I)>	6
Completeness [%]	99.7	95.8	99.9
Redundancy	5	4.5	4.9
CC(1/2)	0.994	0.994	0.447

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		298	250 nL 20 mg/mL AlfC D200A + 250 nL mother liquor (18% w/v PEG3350, 0.1 M Bis-Tris propane, pH 7, 20 mM sodium/potassium phosphate, 1% v/v glycerol), crystals appeared after five days, unliganded crystals were harvested in mother liquor supplemented with 20% v/v glycerol for cryoprotection and 10 mM Fuca(1,6)GlcNAc for ~1 minute, flash-cooled in liquid nitrogen