3KKV

Structure of PKA with a protein Kinase B-selective inhibitor.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Detector technology	CCD
Collection date	2005-07-01
Detector	ADSC QUANTUM 210
Wavelength(s)	1.0000
Spacegroup name	P 21 21 21
Unit cell lengths	72.411, 75.996, 80.390
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	36.206 - 1.800
R-factor	0.2028
Rwork	0.202
R-free	0.22520
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.001
RMSD bond angle	0.590
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	PHENIX ((phenix.refine: 1.5_2))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.860
High resolution limit [Å]	1.800	1.800
Number of reflections	41799
<I/σ(I)>	37
Completeness [%]	99.9	99.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.4	294	Protein stock was at 20mg/ml in 25mM Mes-Bis Tris, pH 6.4, 75mM LiCl, 0.1M EDTA, 1mM DTT. The final concentration of inhibitor in the protein solution was 1mM from a 100mM stock solution of compound dissolved in DMSO.The protein was reacted with inhibitor on ice for 6 hours prior to crystallization.The final concentration of PKI in the protein solution was 2mM from stock 100mM dissolved in water. The final concentration of MEGA8 in the protein solution was 1.5mM from 225mM stock diluted from 790mM in the Hampton additive kit with water. Two microliters of protein were added to 0.35 microliters of 10% glycerol in buffer and put in sitting drops over 15% methanol diluted with water at 4C. Cryo was 100mM MES pH6.4, 100mM LiCl, 25% MeOH, 20% MPD., VAPOR DIFFUSION, SITTING DROP, temperature 294K