6V2A

Complex of double mutant (T89V,K162T) of E. coli L-asparaginase II with L-Asn

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-03-15
Detector	DECTRIS EIGER X 4M
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	151.439, 62.594, 141.659
Unit cell angles	90.00, 117.80, 90.00

Refinement procedure

Resolution	25.980 - 2.000
R-factor	0.1598
Rwork	0.157
R-free	0.20880
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3eca
RMSD bond length	0.014
RMSD bond angle	1.893
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.8.0253)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	40.000	40.000	2.030
High resolution limit [Å]	2.000	5.420	2.000
Rmerge	0.039	0.014	0.509
Rmeas	0.048	0.016	0.633
Rpim	0.027	0.009	0.371
Total number of observations	207458
Number of reflections	73175	4063	3421
<I/σ(I)>	15.5
Completeness [%]	92.6	99	86.7
Redundancy	2.8	3.4	2.6
CC(1/2)		0.999	0.690

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.7	293	Crystals grown at 0.17 M sodium citrate (pH 6) and 17-18% (w/v) PEG3350. Soaked for 1-2 minutes (empirically determined for each crystal) in equivalent solution with 0.025% (w/v) glutaraldehyde. Finally transferred and soaked for 3-5 min in solution containing 40% (w/v) PEG3350, 5 mM L-Asn, and 0.17 M sodium citrate (pH 4.7)