5LCT

Cocrystal structure of cAMP-dependent Protein Kinase (PKA) in complex with a R-methyl-piperazine substituted Fasudil-derivative (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	2016-03-12
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.91841
Spacegroup name	P 21 21 21
Unit cell lengths	59.014, 73.700, 109.837
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.066 - 1.615
R-factor	0.1443
Rwork	0.143
R-free	0.17420
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1q8w
RMSD bond length	0.009
RMSD bond angle	0.963
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]	50.000	1.710
High resolution limit [Å]	1.615	1.615
Rmerge		0.483
Number of reflections	61770
<I/σ(I)>	24.03	3.49
Completeness [%]	99.4	96.4
Redundancy	6.5	6.2

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 (50 mM Stock) Reservoir: 19% Methanol 0.003 mL drop volume, 0.4 mL reservoir volume