8PPE

Human inositol 1,4,5-trisphosphate 3-kinase A (IP3K) catalytic domain in complex with DL-6-deoxy-6-phosphoryloxymethyl-scyllo-inositol 1,2,4-trisphosphate/ADP/Mn

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALBA BEAMLINE XALOC
Synchrotron site	ALBA
Beamline	XALOC
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-03-17
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.979185
Spacegroup name	C 2 2 21
Unit cell lengths	72.554, 97.547, 190.440
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	58.280 - 1.590
R-factor	0.20191
Rwork	0.200
R-free	0.24002
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.006
RMSD bond angle	1.315
Data reduction software	XDS (autoPROC 1.0.5)
Data scaling software	STARANISO (autoPROC 1.0.5, staraniso 2.3.24)
Phasing software	REFMAC
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	58.280	1.660
High resolution limit [Å]	1.590	1.590
Rpim	0.014	0.497
Number of reflections	80256	4014
<I/σ(I)>	22.8	1.3
Completeness [%]	95.4	64.8
Redundancy	13.2	11.1
CC(1/2)	1.000	0.566

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	0.81 M sodium citrate, 0.1M Tris pH 8.5 and 0.1 M NaCl. Protein:precipitant ratio 1:1. Protein concentration: 17 mg/ml. Protein buffer: 20 mM Tris pH 7.5, 50 mM ammonium sulfate and 2 mM DTT. Soaking overnight with 0.83 M sodium citrate, 0.1 M Tris pH 8.5, 0.1 M NaCl, 30 mM ligand, 3 mM ADP and 3 mM MnCl2. Cryoprotectant: Glycerol 30%.