7PIE

Protein kinase A catalytic subunit in complex with PKI5-24 and EN068

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-04-21
Detector	DECTRIS EIGER2 X 16M
Wavelength(s)	1.033193
Spacegroup name	P 21 21 21
Unit cell lengths	58.764, 73.006, 109.486
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.777 - 1.427
R-factor	0.136797965222
Rwork	0.136
R-free	0.15791
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5m6y
RMSD bond length	0.010
RMSD bond angle	1.090
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.777	1.510
High resolution limit [Å]	1.427	1.427
Number of reflections	88126	14074
<I/σ(I)>	28.94	5.47
Completeness [%]	99.7	99.6
Redundancy	13.14	13.36
CC(1/2)	0.999	0.958

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277.15	Protein at 8 mg/mL in 100 mM Mes-Bis-Tris pH 6.9, 75 mM Lithium chloride, 1 mM DTT, 100 mM Sodium-EDTA. Master-Mix: 540 uL of protein + 15 uL 10 mM Mega8 in Water + 45 uL 10 mM PKI5-24 in protein-buffer. 70 % Master-Mix mixed with 30 % 50 mM ligand in DMSO before setting of drops. 15-24 % Methanol/Water in Reservoir.