5MNI

Escherichia coli AGPase mutant R130A apo form

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-09-21
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	94.224, 147.603, 125.572
Unit cell angles	90.00, 91.52, 90.00

Refinement procedure

Resolution	73.800 - 3.090
R-factor	0.2373
Rwork	0.235
R-free	0.27270
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	E coli AGPase tetramer
RMSD bond length	0.005
RMSD bond angle	0.682
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHASER
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	73.800	3.201
High resolution limit [Å]	3.090	3.090
Rmerge	0.085	0.944
Number of reflections	62893
<I/σ(I)>	8.86	1.23
Completeness [%]	99.0	100
Redundancy	3.3	3.4
CC(1/2)	0.997	0.524

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	291	Sitting drop 96 MRC2 well plates using a mosquito crystal robot (TTP Labtech). Crystals were obtained by mixing 0.25 ul of E coli AGPase_R130A at 6.3 mg/ml in 50 mM Tris-HCl pH 7.5, 100 mM NaCl with 0.25 ul of mother liquor containing 14% polyethylene glycol 3.350, 140 mM magnesium formate, 30% ethylene glycol. Crystals grew in 13 days and were frozen under liquid nitrogen.