9BJQ

X-ray crystal structure of wild-type Thermothelomyces thermophilus polysaccharide monooxygenase 9E

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.3
Synchrotron site	ALS
Beamline	5.0.3
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-04-30
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.97648
Spacegroup name	C 2 2 21
Unit cell lengths	99.449, 121.410, 91.909
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.770 - 1.800
R-factor	0.1565
Rwork	0.155
R-free	0.18590
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	0.828
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.770	1.840
High resolution limit [Å]	1.800	1.800
Rmerge	0.070	0.615
Rmeas	0.076	0.668
Rpim	0.029	0.258
Number of reflections	51698	2985
<I/σ(I)>	18.8	3.1
Completeness [%]	99.6	96.6
Redundancy	6.6	6.5
CC(1/2)	0.999	0.870

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	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). 1 ul 30 mg/mL Mt PMO9E in 50 mM MOPS, 50 mM MES was mixed with 1 uL with 0.2 M Ammonium acetate, 0.1 M HEPES pH 7.5, and 30 % PEG 3,350. The crystal was subsequently soaked for 5 minutes in well solution with a final concentration of 1 mM CuSO4. The crystal was 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