2EUV
Principles of protein-DNA recognition revealed in the structural analysis of Ndt80-MSE DNA complexes
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 8.3.1 |
| Synchrotron site | ALS |
| Beamline | 8.3.1 |
| Temperature [K] | 105 |
| Detector technology | CCD |
| Collection date | 2003-07-14 |
| Detector | ADSC QUANTUM 210 |
| Wavelength(s) | 1.072158 |
| Spacegroup name | C 2 2 21 |
| Unit cell lengths | 69.226, 79.248, 160.881 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 16.930 - 1.940 |
| R-factor | 0.195 |
| Rwork | 0.193 |
| R-free | 0.23300 |
| Structure solution method | FOURIER SYNTHESIS |
| Starting model (for MR) | PDB 1MNN - MINUS DNA AT SEQUENCE CHANGES AND 1 BASEPAIR ADJACENT TO THOSE CHANGES |
| RMSD bond length | 0.011 |
| RMSD bond angle | 1.453 |
| Data reduction software | MOSFLM |
| Data scaling software | SCALA |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 38.470 | 2.000 |
| High resolution limit [Å] | 1.900 | 1.900 |
| Rmerge | 0.067 | 0.526 |
| Number of reflections | 35256 | |
| <I/σ(I)> | 7.2 | 1.2 |
| Completeness [%] | 99.9 | 99.9 |
| Redundancy | 4.1 | 3.7 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 7 | 295 | 30% PEG 400, 50 mM bis-tris-propane pH 7.0, 100 mM NaCl, 50 mM CaCl2, and 2 mM DTT. 1:1 Molar ratio protein:DNA, protein at 20mg/ml, VAPOR DIFFUSION, HANGING DROP, temperature 295K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | PEG 400 | ||
| 2 | 1 | 1 | bis-tris-propane | ||
| 3 | 1 | 1 | NaCl | ||
| 4 | 1 | 1 | CaCl2 | ||
| 5 | 1 | 1 | DTT | ||
| 6 | 1 | 1 | H2O | ||
| 7 | 1 | 2 | PEG 400 | ||
| 8 | 1 | 2 | NaCl | ||
| 9 | 1 | 2 | CaCl2 |
