8U8R

Y229F/V290N/S292F Streptomyces coelicolor Laccase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-12-18
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.00003
Spacegroup name	P 43 21 2
Unit cell lengths	177.355, 177.355, 176.956
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	62.700 - 2.860
R-factor	0.183
Rwork	0.181
R-free	0.21370
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	0.938
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHENIX (1.20.1_4487)
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	72.380	2.962
High resolution limit [Å]	2.860	2.860
Rmerge	0.283	1.889
Number of reflections	65563	6445
<I/σ(I)>	10.32
Completeness [%]	100.0
Redundancy	13.4
CC(1/2)	0.992	0.607

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		296	Crystals were prepared using hanging drop vapor-diffusion technique at room temperature (~296 K). Protein is at a concentration of 18.5 mg/ml in 50 mM H3BO3, 0.1 M NaCl, pH 9.0 buffer. The well buffer contains 0.1 M glycine, 0.3-0.6 M NaCl, pH 9.0, and 37-39% (v/v) PEG (polyethylene glycol) monomethyl ether 550. 500 uL of well buffer is added to each well and protein is mixed with well buffer at a 1.5 uL:1.5 uL ratio. The crystal growth time was ca. 1-2 weeks