8GM5

Functional construct of the Eukaryotic elongation factor 2 kinase bound to Calmodulin, ADP and to the A-484954 inhibitor and showing two conformations for the 498-520 loop

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-2
Synchrotron site	NSLS-II
Beamline	17-ID-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-05-28
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.97934
Spacegroup name	P 1 21 1
Unit cell lengths	80.909, 61.041, 88.958
Unit cell angles	90.00, 111.26, 90.00

Refinement procedure

Resolution	45.940 - 2.120
R-factor	0.183
Rwork	0.181
R-free	0.22500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	7shq
RMSD bond length	0.003
RMSD bond angle	0.518
Data reduction software	XDS (Jan 10, 2022)
Data scaling software	Aimless (0.7.7)
Phasing software	PHASER (2.3.90)
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	82.906	82.906	2.338
High resolution limit [Å]	2.120	6.673	2.124
Rmerge	0.122	0.049	0.889
Rmeas	0.137	0.054	1.009
Rpim	0.060	0.024	0.474
Total number of observations		7601	6853
Number of reflections	30789	1539	1539
<I/σ(I)>	5.89	13.44	1.46
Completeness [%]	92.0	99	53.4
Completeness (spherical) [%]		99.0	13.5
Completeness (ellipsoidal) [%]		99.0	53.4
Redundancy	4.93	4.94	4.45
CC(1/2)	0.996	0.997	0.567
Anomalous completeness (spherical)		98.7	13.8
Anomalous completeness		98.7	54.1
Anomalous redundancy		2.7	2.3
CC(ano)		-0.444	0.030
|DANO|/σ(DANO)		0.4	0.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	6.5	295.15	Cocktail:16.55% PEG-3350, 0.2 M NaF, 100 mM BisTris-Propane Protein solution: 10.3 mg/mL 20 mM Tris pH 7.5, 100 mM NaCl, 3 mM CaCl2, 1mM TCEP, 1.5 m Inhibitor , 3.1 % DMSO 2protein/1cocktail (0.2 ul total)