6V2B

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	2017-02-16
Detector	DECTRIS EIGER X 4M
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	151.224, 62.578, 140.886
Unit cell angles	90.00, 117.67, 90.00

Refinement procedure

Resolution	26.550 - 2.050
R-factor	0.1547
Rwork	0.152
R-free	0.20130
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3eca
RMSD bond length	0.015
RMSD bond angle	1.915
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	2.090
High resolution limit [Å]	2.050	5.560	2.050
Rmerge	0.047	0.014	0.512
Rmeas	0.057	0.017	0.632
Rpim	0.033	0.009	0.364
Total number of observations	199695
Number of reflections	70596	3818	3449
<I/σ(I)>	12.5
Completeness [%]	95.7	99.1	94.6
Redundancy	2.8	3.4	2.5
CC(1/2)		0.999	0.706

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	298	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 10-20 sec in solution containing 40% (w/v) PEG3350, 5 mM L-Asn, and 0.17 M sodium citrate (pH 5.5)