5M0B

Cocrystal structure of cAMP-dependent Protein Kinase (PKA) in complex with a short-chained N-(2-aminoethyl)isoquinoline-5-sulfonamide) Fasudil-derivative (Ligand 03)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM14
Synchrotron site	ESRF
Beamline	BM14
Temperature [K]	100
Detector technology	CCD
Collection date	2015-04-13
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97625
Spacegroup name	P 21 21 21
Unit cell lengths	57.945, 71.736, 109.922
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.876 - 1.506
R-factor	0.1334
Rwork	0.132
R-free	0.15940
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1q8w
RMSD bond length	0.008
RMSD bond angle	0.955
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]	45.080	1.600
High resolution limit [Å]	1.506	1.506
Rmerge		0.498
Number of reflections	73047
<I/σ(I)>	26.28	3.89
Completeness [%]	99.9	99.7
Redundancy	7.32	7.26

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: 18% Methanol 0.003 mL drop volume, 0.4 mL reservoir volume