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) |