7PNS

Protein kinase A catalytic subunit in complex with PKI5-24 and EN081

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	69.444, 73.122, 77.172
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.171 - 1.855
R-factor	0.17557903644
Rwork	0.174
R-free	0.20153
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5m6y
RMSD bond length	0.010
RMSD bond angle	1.006
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]	42.171	1.960
High resolution limit [Å]	1.855	1.855
Number of reflections	33232	5242
<I/σ(I)>	28.79	5.88
Completeness [%]	97.0	96.1
Redundancy	13.31	13.58
CC(1/2)	0.999	0.962

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.