9QV0

1-Phosphofructokinase (FruK) from E. coli with bound fructose 1-phosphate and ADP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-11-20
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9763
Spacegroup name	I 2 2 2
Unit cell lengths	63.150, 98.460, 140.620
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.760 - 1.550
R-factor	0.1711
Rwork	0.171
R-free	0.17980
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.774
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]	19.760	19.760	1.590
High resolution limit [Å]	1.550	6.930	1.550
Rmerge	0.062	0.023	1.123
Rmeas	0.066	0.024	1.199
Number of reflections	62272	765	4501
<I/σ(I)>	22.46		2.5
Completeness [%]	97.5
Redundancy	8.7
CC(1/2)	1.000	0.999	0.830

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	FruK (3.4 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, 2 mM fructose-1-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 35% (v/v).