6UWJ

Intermediate engineered variant of I-OnuI meganuclease with improved thermostability and partially altered specificity

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	2019-04-25
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.977
Spacegroup name	P 21 21 21
Unit cell lengths	38.288, 62.586, 157.405
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.985 - 1.852
R-factor	0.1935
Rwork	0.191
R-free	0.23420
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3qqy
RMSD bond length	0.010
RMSD bond angle	1.136
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER (2.8.1)
Refinement software	PHENIX (1.13)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.920
High resolution limit [Å]	1.850	3.990	1.850
Rmerge	0.035	0.027	0.162
Rmeas	0.037	0.028	0.175
Rpim	0.011	0.008	0.063
Total number of observations	374757
Number of reflections	33122	3578	3166
<I/σ(I)>	20.2
Completeness [%]	99.7	99.9	96.7
Redundancy	11.3	11.6	7.3
CC(1/2)		0.999	0.988

Crystallization Conditions

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