10JU

Crystal Structure of serine/threonine-protein kinase (AEK1) T376D, S395D Mutant from Trypanosoma brucei (AMP-PNP)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 19-ID
Synchrotron site	NSLS-II
Beamline	19-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2025-12-13
Detector	DECTRIS EIGER2 XE 9M
Wavelength(s)	0.9786
Spacegroup name	I 2 2 2
Unit cell lengths	86.538, 88.475, 201.132
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.240 - 2.150
R-factor	0.2202
Rwork	0.219
R-free	0.25040
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.005
RMSD bond angle	0.641
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((2.0_5936: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.240	2.220
High resolution limit [Å]	2.150	2.150
Rmerge	0.079	1.693
Rmeas	0.082	1.758
Rpim	0.023	0.473
Total number of observations	564111	49777
Number of reflections	42374	3628
<I/σ(I)>	17.2	1.6
Completeness [%]	100.0
Redundancy	13.3	13.7
CC(1/2)	0.999	0.747

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	291	Index F6 0.2M Ammonium sulfate 0.1M Bis-Tris pH 5.5 25% PEG 3350. TrbrA.01480.a.WW4.PS38793 at 13.5 mg/mL. The C-terminal tail ~60 residues was disordered in each subunit. Residue Ser 71 in subunit A contained a large amount of density near the OG atom. This was modeled as a phosphoserine (SEP) although this is not a predicted phosphorylation site. 2mM AMP-PNP and 2mM MgCl2 added prior to crystallization. Prominent AMP-PNP density in subunit B but disorder in the phosphate portion. plate 20522 F6 drop 2, Puck: PSL-0904, Cryo: 80% crystallant + 20% glycerol