2PHK

THE CRYSTAL STRUCTURE OF A PHOSPHORYLASE KINASE PEPTIDE SUBSTRATE COMPLEX: KINASE SUBSTRATE RECOGNITION

Experimental procedure

Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM02
Synchrotron site	ESRF
Beamline	BM02
Temperature [K]	100
Detector technology	CCD AREA DETECTOR
Collection date	1997-03-11
Detector	XRII
Spacegroup name	P 32 2 1
Unit cell lengths	65.300, 65.300, 145.800
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.000 - 2.600
R-factor	0.253
Rwork	0.236
R-free	0.30000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phk
RMSD bond length	0.015 *
RMSD bond angle	1.800 *
Data reduction software	MOSFLM
Data scaling software	CCP4
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.740
High resolution limit [Å]	2.600	2.600
Rmerge	0.098	0.360
Total number of observations	21644 *
Number of reflections	10062
<I/σ(I)>	7.1	1.3
Completeness [%]	86.3	78.1
Redundancy	2.15	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	8.2 *		pH 6.9

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	2-3 (mg/ml)
10	1	reservoir	PEG8000	5 (%)
11	1	reservoir	HEPES	50 (mM)
12	1	reservoir	glycerol	10 (%(v/v))
13	1	reservoir	dithiothreitol	10 (mM)
14	1	reservoir		0.02 (%(w/v))
15	1	reservoir		5 (mM)
2	1	drop	MC-peptide	10 (mM)
3	1	drop	AMPPNP	3 (mM)
4	1	drop	HEPES/NaOH	10 (mM)
5	1	drop	glycerol	2 (%(v/v))
6	1	drop	dithiothreitol	10 (mM)
7	1	drop		0.2 (%(w/v))
8	1	drop	EDTA	0.1 (mM)
9	1	drop		5 (mM)