9QV3

1-Phosphofructokinase mutant K95T/G110S from E. coli with bound fructose 1-phosphate and ADP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-02-12
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.976254
Spacegroup name	I 2 2 2
Unit cell lengths	62.690, 98.900, 139.590
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.620 - 1.410
R-factor	0.1601
Rwork	0.160
R-free	0.17540
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.012
RMSD bond angle	1.127
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX ((1.21.2_5419: ???))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	19.620	19.620	1.450
High resolution limit [Å]	1.410	6.310	1.410
Rmerge	0.075	0.061	0.605
Rmeas	0.082	0.068	0.660
Number of reflections	83528	1005	6126
<I/σ(I)>	12.85
Completeness [%]	99.8
Redundancy	6.26
CC(1/2)	0.995	0.986	0.855

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	289	The FruK variant K95T/G110S (3.5 mg/mL) in 20 mM MOPS/KOH buffer (pH 7.4) containing 75 mM KCl and 1 M L-proline was mixed in a 1:1 ratio with condition containing 1 M ammonium sulfate, 75 mM TRIS/HCl (pH 8.5), 150 mM lithium sulfate, 8 mM fructose-6-phosphate, 4 mM ADP, and 10 mM magnesium chloride. The final drop volume was 0.8 microliters. Before plunge freezing the crystals in liquid nitrogen, glycerol was added to the drop as a cryo-protectant to a final concentration of 30% (v/v).