9NJJ

F195L/I200F/M298L Streptomyces coelicolor Laccase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-10-28
Detector	DECTRIS EIGER2 S 9M
Wavelength(s)	1.000006
Spacegroup name	P 43 3 2
Unit cell lengths	177.832, 177.832, 177.832
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	79.530 - 2.640
R-factor	0.1867
Rwork	0.185
R-free	0.20770
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.477
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]	79.530	2.734
High resolution limit [Å]	2.640	2.640
Rmerge	0.272	2.903
Rmeas	0.275	2.940
Number of reflections	28814	2808
<I/σ(I)>	16.58	1.69
Completeness [%]	100.0	99.93
Redundancy	39.5	39.8
CC(1/2)	0.998	0.849

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	9	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, 0.5 M NaCl, pH 9.0, and 37-40% (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 week.