6V27

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	2019-02-17
Detector	DECTRIS EIGER X 4M
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	151.302, 62.412, 141.753
Unit cell angles	90.00, 117.85, 90.00

Refinement procedure

Resolution	26.500 - 2.300
R-factor	0.1643
Rwork	0.160
R-free	0.25130
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3eca
RMSD bond length	0.015
RMSD bond angle	2.070
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	2.340
High resolution limit [Å]	2.300	6.240	2.300
Rmerge	0.073	0.022	0.513
Rmeas	0.091	0.027	0.640
Rpim	0.053	0.015	0.377
Total number of observations	138795
Number of reflections	51978	2675	2587
<I/σ(I)>	9.9
Completeness [%]	98.9	97.5	98.3
Redundancy	2.7	2.9	2.5
CC(1/2)		0.998	0.722

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 10-20 sec in solution containing 40% (w/v) PEG3350, 5 mM L-L-Asp, and 0.17 M sodium citrate (pH 4.7)