1GKZ
Branched-chain alpha-ketoacid dehydrogenase kinase (BCK) complxed with ADP
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | APS BEAMLINE 19-BM |
| Synchrotron site | APS |
| Beamline | 19-BM |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2001-02-15 |
| Detector | SBC-CAT |
| Spacegroup name | P 42 21 2 |
| Unit cell lengths | 128.917, 128.917, 74.105 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 40.770 - 2.200 |
| R-factor | 0.225 * |
| Rwork | 0.237 |
| R-free | 0.28500 * |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | BRANCHED-CHAIN ALPHA-KETOACID DEHYDROGENASE KINASE (BCK) COMLEXED WITH ATP-GAMMA-S |
| RMSD bond length | 0.012 * |
| RMSD bond angle | 1.700 * |
| 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] | 40.770 | 2.240 |
| High resolution limit [Å] | 2.200 | 2.290 * |
| Rmerge | 0.044 * | 0.606 * |
| Number of reflections | 32115 | |
| <I/σ(I)> | 41.7 | 2.7 |
| Completeness [%] | 95.7 * | 85.3 * |
| Redundancy | 4.5 * | 3.2 * |
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 | Mg-ADP | ||
| 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) |
