6UWG

Engineered variant of I-OnuI meganuclease with improved thermostability and E178D mutation at catalytic site

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	108
Detector technology	CCD
Collection date	2019-10-04
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	39.324, 73.516, 163.526
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.430 - 2.220
R-factor	0.197
Rwork	0.193
R-free	0.24060
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	2.300
High resolution limit [Å]	2.220	4.780	2.220
Rmerge	0.057	0.033	0.341
Rmeas	0.062	0.036	0.381
Rpim	0.024	0.014	0.166
Total number of observations	158867
Number of reflections	24263	2619	2307
<I/σ(I)>	11.4
Completeness [%]	99.5	98.9	95.8
Redundancy	6.5	6.7	5
CC(1/2)		0.999	0.909

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	200mM Calcium acetate, 100mM sodium acetate pH 5.5, 100mM sodium acetate pH 6.0, 35% PEG 400