2Q2U
Structure of Chlorella virus DNA ligase-product DNA complex
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | NSLS BEAMLINE X9A |
| Synchrotron site | NSLS |
| Beamline | X9A |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2006-05-05 |
| Detector | MAR CCD 165 mm |
| Wavelength(s) | 0.9795 |
| Spacegroup name | P 1 |
| Unit cell lengths | 66.475, 92.895, 89.633 |
| Unit cell angles | 70.32, 78.45, 89.87 |
Refinement procedure
| Resolution | 24.880 - 3.000 |
| Rwork | 0.249 |
| R-free | 0.28300 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | Structure of Chlorella virus DNA ligase bound to nicked DNA |
| RMSD bond length | 0.007 |
| RMSD bond angle | 1.100 |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | AMoRE |
| Refinement software | CNS (1.1) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 50.000 | 3.110 |
| High resolution limit [Å] | 3.000 | 3.000 |
| Rmerge | 0.078 | 0.455 |
| Number of reflections | 39849 | |
| <I/σ(I)> | 7.3 | 1.4 |
| Completeness [%] | 95.8 | 87.7 |
| Redundancy | 1.9 | 1.7 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 6.5 | 295 | A mixture of ChVLig (230 microM), nicked duplex DNA (220 microM) and 2 mM EDTA was added to an equal volume of a well solution containing 100 mM Bis-Tris-HCl (pH 6.5), 30 mM ammonium acetate, 22% PEG-4000. Crystals were grown at 22 C by the sitting-drop vapor diffusion method. Crystals appeared after 3 days. Crystals of ChVLig in complex with nicked DNA were transferred to a solution containing 100 mM Bis-Tris-HCl (pH 6.5), 110 mM ammonium acetate, 22.5% PEG-4000, 5 mM MnCl2 for 5 min, then placed into a solution containing the same components plus 15% glycerol prior to flash-freezing the crystals in liquid nitrogen., VAPOR DIFFUSION, SITTING DROP, temperature 295K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | Bis-Tris-HCL | ||
| 10 | 1 | 3 | ammonium acetate | ||
| 11 | 1 | 3 | glycerol | ||
| 2 | 1 | 1 | EDTA | ||
| 3 | 1 | 1 | PEG-4000 | ||
| 4 | 1 | 1 | ammonium acetate | ||
| 5 | 1 | 2 | EDTA | ||
| 6 | 1 | 2 | PEG-4000 | ||
| 7 | 1 | 2 | ammonium acetate | ||
| 8 | 1 | 3 | EDTA | ||
| 9 | 1 | 3 | PEG-4000 |
