9NJI

M298L Streptomyces coelicolor Laccase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-04-11
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.0330
Spacegroup name	P 43 3 2
Unit cell lengths	177.228, 177.228, 177.228
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.310 - 2.600
R-factor	0.1864
Rwork	0.186
R-free	0.20160
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.484
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]	44.310	2.693
High resolution limit [Å]	2.600	2.600
Rmerge	0.288	2.191
Rmeas	0.292	2.218
Number of reflections	29840	2930
<I/σ(I)>	13.83	1.76
Completeness [%]	100.0	100
Redundancy	40.5	41.3
CC(1/2)	0.997	0.773

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.