6V2M

Structure of Escherichia coli Asp269Asn mutant phosphoenolpyruvate carboxykinase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CLSI BEAMLINE 08B1-1
Synchrotron site	CLSI
Beamline	08B1-1
Temperature [K]	105
Detector technology	CCD
Collection date	2014-02-28
Detector	RAYONIX MX300HE
Wavelength(s)	0.98
Spacegroup name	C 1 2 1
Unit cell lengths	124.780, 94.220, 46.540
Unit cell angles	90.00, 94.79, 90.00

Refinement procedure

Resolution	46.380 - 1.660
Rwork	0.185
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ayl
Data reduction software	XDS
Data scaling software	XDS
Phasing software	MOLREP
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.110	1.719
High resolution limit [Å]	1.660	1.660
Rmerge	0.115	2.915
Rmeas	0.132	3.340
Rpim	0.064	1.613
Number of reflections	63107	6242
<I/σ(I)>	7.96	0.65
Completeness [%]	99.7	98.71
Redundancy	4.2	4.2
CC(1/2)	0.997	0.175

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH		293	2ul drop with 2 mg/ml protein, 5 mM Calcium chloride, 5mM Magnesium chloride, 2 mM ATP, 2mM Pyruvate, 1 mM EDTA, 200 mM Ammonium acetate, 100 mM Sodium acetate, 0.01 mM DTT, 30% PEG 4000 was added to 2 ul drop containing 2 M sodium aceate, 0.1 M Tris pH 8.2 30% PEG 4000. After a week a rod like crystal was removed and soaked in a solution with 30% glycerol 1mM EDTA, 100 mM sodium acetate 200mM ammonium acetate and 12% PEG 4000. The crystal was put into a loop and flash cooled in liquid notrogen