1GKX

Branched-chain alpha-ketoacid dehydrogenase kinase (BCK)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2001-06-15
Detector	SBC-CAT
Spacegroup name	P 42 21 2
Unit cell lengths	129.370, 129.370, 73.350
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.500 * - 2.980 *
R-factor	0.243 *
Rwork	0.241
R-free	0.27200 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	BRANCHED-CHAIN ALPHA-KETOACID DEHYDROGENASE KINASE (BCK) COMPLEXED WITH ATP-GAMMA-S
RMSD bond length	0.010 *
RMSD bond angle	1.500 *
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	CNS (1.0)
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.500 *	2.340
High resolution limit [Å]	2.980 *	2.980 *
Rmerge	0.023 *	0.785 *
Number of reflections	27710
<I/σ(I)>	29	1.8
Completeness [%]	99.4 *	94.5 *
Redundancy	8.2 *	7.6 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	7.5	20 *	VAPOR DIFFUSION, 20C, EQUAL AMOUNTS OF BCK (20 MG/ML IN 50 MM HEPES, PH 7.5, 1 M SODIUM CHLORIDE, 250 MM POTASSIUM CHLORIDE, 300 MM ARGININE, 20 MM BETA- MERCAPTOETHANOL, 2 MM BENZAMIDINE, 2 MM MAGNESIUM CHLORIDE, 0.5 MM PMSF, 10% (W/V) GLYCEROL) AND RESERVOIR (8%(W/V) PEG-6000, 5% (V/V) ETHYLENE GLYCOL, 1 M NACL, 20 MM BETA-MERCAPTOETHANOL)

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20 (mg/ml)
10	1	drop	glycerol	10 (%(v/v))
11	1	reservoir	PEG6000	8 (%(w/v))
12	1	reservoir	ethylene glycol	5 (%(v/v))
13	1	reservoir		1 (M)
14	1	reservoir	beta-mercaptoethanol	20 (mM)
2	1	drop	HEPES	50 (mM)
3	1	drop		1 (M)
4	1	drop		250 (mM)
5	1	drop	arginine	300 (mM)
6	1	drop	beta-mercaptoethanol	20 (mM)
7	1	drop	benzamidine	1 (mM)
8	1	drop		2 (mM)
9	1	drop	PMSF	0.5 (mM)