3ERC
Crystal structure of the heterodimeric vaccinia virus mRNA polyadenylate polymerase with three fragments of RNA and 3'-deoxy ATP
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | SSRL BEAMLINE BL7-1 |
| Synchrotron site | SSRL |
| Beamline | BL7-1 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2008-07-27 |
| Detector | ADSC QUANTUM 315 |
| Wavelength(s) | 0.97670 |
| Spacegroup name | P 1 |
| Unit cell lengths | 69.925, 77.153, 108.030 |
| Unit cell angles | 89.52, 73.45, 63.76 |
Refinement procedure
| Resolution | 38.550 - 3.210 |
| R-factor | 0.2451 |
| Rwork | 0.245 |
| R-free | 0.30900 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 2ga9 |
| RMSD bond length | 0.012 |
| RMSD bond angle | 1.400 |
| Data reduction software | HKL-2000 |
| Data scaling software | HKL-2000 |
| Phasing software | PHASER |
| Refinement software | CNS (1.2) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 50.000 | 3.410 |
| High resolution limit [Å] | 3.210 | 3.210 |
| Rmerge | 0.107 | 0.427 |
| Number of reflections | 30209 | |
| <I/σ(I)> | 12.67 | 1.93 |
| Completeness [%] | 93.8 | 94.6 |
| Redundancy | 3.2 | 3.2 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 8.7 | 295 | Proteins(4-5mg.ml) in 10mM Tris-HCl, pH8.7, 75 mM NaCl, 0.5 mM DTT, mixed with equal volume buffer which composed of 10mM Tris-HCl, pH 8.7, 15-20% PEG 4000, 5% glycerol, 0.5 mM DTT. Room temperature for several days., VAPOR DIFFUSION, SITTING DROP, temperature 295K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | Tris-HCl | ||
| 2 | 1 | 1 | NaCl | ||
| 3 | 1 | 1 | DTT | ||
| 4 | 1 | 1 | PEG 4000 | ||
| 5 | 1 | 1 | glycerol | ||
| 6 | 1 | 2 | Tris-HCl | ||
| 7 | 1 | 2 | DTT | ||
| 8 | 1 | 2 | PEG 4000 | ||
| 9 | 1 | 2 | glycerol |
