6UW0

Engineered variant of I-OnuI meganuclease with improved thermostability and fully altered specificity targeting cholera toxin A subunit

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.1
Synchrotron site	ALS
Beamline	5.0.1
Temperature [K]	108
Detector technology	PIXEL
Collection date	2019-06-20
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.977
Spacegroup name	P 21 21 21
Unit cell lengths	75.246, 79.679, 168.941
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.990 - 2.720
R-factor	0.2276
Rwork	0.223
R-free	0.28090
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3qqy
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER (2.8.1)
Refinement software	PHENIX (1.17_3644)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.800
High resolution limit [Å]	2.700	5.810	2.700
Rmerge	0.075	0.035	0.416
Rmeas	0.078	0.037	0.439
Rpim	0.023	0.011	0.138
Total number of observations	315424
Number of reflections	27693	3004	2493
<I/σ(I)>	8.4
Completeness [%]	98.7	100	90.5
Redundancy	11.4	11.3	9.1
CC(1/2)		1.000	0.965

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	293	100mM HEPES pH 8.5, 200mM ammonium sulfate, 30% PEG 3350