5MQ5

A protease-resistant N24S Escherichia coli Asparaginase mutant with outstanding stability and enhanced anti-leukaemic activity

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-06-26
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.976251
Spacegroup name	C 1 2 1
Unit cell lengths	152.003, 62.394, 142.271
Unit cell angles	90.00, 117.92, 90.00

Refinement procedure

Resolution	12.970 - 1.600
Rwork	0.147
R-free	0.19790
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3eca
RMSD bond length	0.007
RMSD bond angle	0.794
Data reduction software	iMOSFLM (7.2.1)
Data scaling software	SCALA (CCP4i v7)
Phasing software	PHASER (CCP4i v7)
Refinement software	PHENIX

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.160	1.480
High resolution limit [Å]	1.427	1.430
Rmerge	0.061	1.284
Rmeas	0.076	1.816
Rpim	0.044	1.284
Number of reflections	195240	11874
<I/σ(I)>	8.6
Completeness [%]	90.0	55.9
Redundancy	2.6	1.9
CC(1/2)	0.998	0.233

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	294.15	100 mM 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, 5% w/v PEG 8000, 4% v/v ethylene glycol. Soaking in 0.1 mM L-asparatate.