8R56

Crystal structure of GH31 family Sulfoquinovosidase BmSQase in covalent complex with SQ-aziridine (SQZ)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-09-15
Detector	DECTRIS EIGER2 XE 16M
Wavelength(s)	0.97624
Spacegroup name	P 1 21 1
Unit cell lengths	62.524, 194.726, 117.654
Unit cell angles	90.00, 92.29, 90.00

Refinement procedure

Resolution	62.550 - 2.450
R-factor	0.21347
Rwork	0.212
R-free	0.24405
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.525
Data reduction software	xia2
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0419)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	74.990	2.490
High resolution limit [Å]	2.450	2.450
Rmerge	0.197	1.174
Rmeas	0.212	1.267
Rpim	0.080	0.474
Total number of observations	737059	36181
Number of reflections	102876	5135
<I/σ(I)>	9.3	2.2
Completeness [%]	99.9
Redundancy	7.2	7
CC(1/2)	0.986	0.637

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION		293	A crystal of BmSQase was grown using a 30 mg mL-1 protein solution in buffer C, 0.15 uL protein. 0.15 uL mother liquor, the latter comprising 0.2 M KSCN and 20% w/v PEG (polyethylene glycol) 3350 in 0.1 M Bis-TRIS propane buffer pH 7.5. A stock solution of 1 was prepared at 10 mM in mother liquor. Crystals were soaked for 5 min with 0.3 ul of 1 to a final concentration of 3 mM.