9D0Z

X-ray crystal structure of H157Q variant Thermothelomyces thermophilus polysaccharide monooxygenase 9E

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	2023-12-15
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.000046
Spacegroup name	C 2 2 21
Unit cell lengths	102.410, 122.473, 91.478
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.680 - 1.800
R-factor	0.2125
Rwork	0.211
R-free	0.23630
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.848
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.21.1_5286)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.740	1.840
High resolution limit [Å]	1.800	1.800
Rmerge	0.135	0.537
Rmeas	0.157	0.627
Rpim	0.080	0.377
Number of reflections	53154	3123
<I/σ(I)>	6	2
Completeness [%]	99.5	99.7
Redundancy	3.6	3.8
CC(1/2)	0.985	0.811

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	295	The crystal was grown by hanging drop in conditions optimized from Hampton Index HT G8 (0.2 M Ammonium acetate, 0.1 M HEPES pH 7.5, 25 % w/v polyethylene glycol 3,350). H157Q Mt PMO9E was first reconstituted with excess copper and buffer exchanged to buffer with 50 mM MOPS and 50 mM HEPES pH 6. 1 ul 30 mg/mL H157Q Mt PMO9E was then mixed with 1 uL with 0.2 M Ammonium acetate, 0.1 M HEPES pH 7.5, and 26 % PEG 3,350 in a hanging drop. Crystals grew within 4 days. The crystal was looped with a 0.1-0.2 mm loop, then cryoprotected for 10 seconds in well solution mixed with cryoprotectants to a final concentration of 8% w/v sucrose, 2% w/v glucose, 8% v/v glycerol, and 8% ethylene glycol