6YPP

Crystal structure of the cAMP-dependent protein kinase A cocrystallized with PKI (5-24). Soaking of aminofasudil and displacing it with the fragment isoquinoline.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.1
Synchrotron site	BESSY
Beamline	14.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-06-02
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.91840
Spacegroup name	P 21 21 21
Unit cell lengths	52.499, 71.658, 97.779
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.890 - 1.750
R-factor	0.1787
Rwork	0.177
R-free	0.20290
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6f14
RMSD bond length	0.009
RMSD bond angle	0.910
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.890	1.850
High resolution limit [Å]	1.747	1.750
Number of reflections	37708	5905
<I/σ(I)>	20.67	2.56
Completeness [%]	98.9
Redundancy	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.9	277	0,2 mM PKA in 100 mM MES-BIS-Tris-Buffer, 1 mM dithiothreitol, 0.1 mM sodium EDTA, 75 mM LiCl, 0.2 mM Mega 8 and 23 % methanol (v/v) 0.003 mL drop volume, 0.5 mL reservoir volume Soaking of aminofasudil: in buffer with 10% of ligand in DMSO (50 mM stock) and 30% MPD Displacement with isoquinoline: in buffer with 10% of ligand in DMSO and 30% MPD