5M0L

Cocrystal structure of cAMP-dependent Protein Kinase (PKA) in complex with the methylated Fasudil-derived fragment N-methylisoquinoline-5-sulfonamide (Ligand 02)

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	2014-11-22
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.91841
Spacegroup name	P 21 21 21
Unit cell lengths	65.814, 79.707, 84.511
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	32.907 - 1.465
R-factor	0.1535
Rwork	0.152
R-free	0.17280
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1q8w
RMSD bond length	0.008
RMSD bond angle	1.046
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]	34.090	1.550
High resolution limit [Å]	1.465	1.465
Rmerge		0.495
Number of reflections	75918
<I/σ(I)>	19.11	2.7
Completeness [%]	98.7	97.4
Redundancy	4.07	4.18

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.9	277.15	Drop: 10 mg/ml PKA (0.240 mM) 30 mM MBT (MES/Bis-Tris Puffer pH 6.9) 1 mM DTT 0.1 mM EDTA 75 mM LiCl 0.03 mM Mega 8 0.07mM PKI (Sigma: P7739) 1.2 mM ligand solved in DMSO (100 mM Stock) Reservoir: 16% Methanol 0.003 mL drop volume, 0.4 mL reservoir volume