6V29

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-05-22
Detector	DECTRIS EIGER X 4M
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	151.616, 62.430, 143.066
Unit cell angles	90.00, 118.09, 90.00

Refinement procedure

Resolution	26.000 - 2.000
R-factor	0.1454
Rwork	0.142
R-free	0.20370
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3eca
RMSD bond length	0.020
RMSD bond angle	1.922
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.8.0158)

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	0.060	0.011	0.526
Rmeas	0.072	0.013	0.639
Rpim	0.039	0.007	0.358
Total number of observations	256387
Number of reflections	79279	4099	3866
<I/σ(I)>	10.7
Completeness [%]	99.1	98.8	98.3
Redundancy	3.2	3.5	2.9
CC(1/2)		1.000	0.717

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.2	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 10-20 sec in solution containing 40% (w/v) PEG3350, 5 mM L-L-Asp, and 0.17 M sodium citrate (pH 6.2)