1O6K

Structure of activated form of PKB kinase domain S474D with GSK3 peptide and AMP-PNP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2002-07-15
Detector	ADSC CCD
Spacegroup name	P 21 21 21
Unit cell lengths	44.906, 60.998, 129.410
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 * - 1.700
R-factor	0.205
Rwork	0.205
R-free	0.23400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	CDK1
RMSD bond length	0.012
RMSD bond angle	1.470 *
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.800
High resolution limit [Å]	1.700	1.700
Rmerge	0.078	0.186
Total number of observations	116971 *
Number of reflections	27820 *
<I/σ(I)>	6	2.3
Completeness [%]	82.4	63.7
Redundancy	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	unknown *	7.5	20 *	10 MG/ML PROTEIN 20 PEG 4K, 10% ISOPROPONAL, 5 MM DTT, pH 7.50

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	drop	AMP-PNP-MnCl2	5 (mM)
3	1	drop	GSK3beta-peptide	0.6 (mM)
4	1	reservoir	PEG4000	20 (%(w/v))
5	1	reservoir	iso-propanol	10 (%(v/v))
6	1	reservoir	HEPES	0.1 (M)	pH7.5
7	1	reservoir	dithiothreitol	5 (mM)