1GJV

Branched-chain alpha-ketoacid dehydrogenase kinase (BCK) complxed with ATP-gamma-S

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2001-04-15
Detector	MSC
Spacegroup name	P 42 21 2
Unit cell lengths	127.722, 127.722, 74.314
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	36.940 - 2.700
R-factor	0.231 *
Rwork	0.218
R-free	0.26900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	BRANCHED-CHAIN ALPHA-KETOACID DEHYDROGENASE KINASE (BCK) SELENOMETHIONINE VARIANT
RMSD bond length	0.011
RMSD bond angle	22.100 *
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	CNS (1.0)
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	36.900	2.800
High resolution limit [Å]	2.700	2.700
Rmerge	0.061 *	0.582 *
Number of reflections	17401
<I/σ(I)>	24.4	2
Completeness [%]	99.6	96.8
Redundancy	4.6	3.6

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 MG-ATPGAMMAS, 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	drop	ATP gammaS
12	1	reservoir	PEG6000	8 (%(w/v))
13	1	reservoir	ethylene glycol	5 (%(v/v))
14	1	reservoir		1 (M)
15	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)