8PPA

Human inositol 1,4,5-trisphosphate 3-kinase A (IP3K) catalytic domain in complex with D-myo-inositol 1,4,6-trisphosphate/AMP-PNP/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	2021-06-19
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.979264
Spacegroup name	C 2 2 21
Unit cell lengths	72.653, 97.919, 192.066
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.910 - 1.730
R-factor	0.19464
Rwork	0.193
R-free	0.21823
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.006
RMSD bond angle	1.402
Data reduction software	XDS
Data scaling software	XDS
Phasing software	REFMAC
Refinement software	REFMAC (v5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.860	1.760
High resolution limit [Å]	1.730	1.730
Rpim	0.037	0.417
Number of reflections	64275	3876
<I/σ(I)>	12.1	1.8
Completeness [%]	89.5	100
Redundancy	11.8	12.2
CC(1/2)	0.994	0.756

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: 18 mg/ml. Protein buffer: 20 mM Tris pH 7.5, 50 mM ammonium sulfate and 2 mM DTT. Soaking 2h with 1.5 M lithium sulfate, 0.1 M Tris pH 8.5, 5 mM ligand, 3 mM AMP-PNP and 3 mM MnCl2.