7S3V

Structure of HsKYNase_66, an evolved variant of human kynureninase with greatly increased activity towards kynurenine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-D
Synchrotron site	APS
Beamline	23-ID-D
Temperature [K]	93
Detector technology	PIXEL
Collection date	2017-11-15
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.03322
Spacegroup name	I 41 2 2
Unit cell lengths	140.890, 140.890, 286.371
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.344 - 3.249
R-factor	0.1896
Rwork	0.185
R-free	0.22780
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2hzp
RMSD bond length	0.004
RMSD bond angle	0.732
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.11.1_2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	3.310
High resolution limit [Å]	3.249	8.810	3.250
Rmerge	0.164	0.056	0.791
Rmeas	0.191	0.065	0.921
Rpim	0.095	0.032	0.460
Number of reflections	21474	1079	1064
<I/σ(I)>	4.3
Completeness [%]	92.7	84.2	94.7
Redundancy	3.6	3.5	3.6
CC(1/2)		0.992	0.574

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277	8% PEG8000, 100 mM imidazole, 50 mM MgCl2, 0.2 mM PLP, 5% sucrose (diffracting crystal proteins were briefly supplemented with 0.005 mg/mL trypsin prior to sitting drop vapor diffusion in order to remove flexible terminal ends).