2ETW
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 | 70.299, 78.919, 161.625 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 23.570 - 1.670 |
| R-factor | 0.181 |
| Rwork | 0.180 |
| R-free | 0.19900 |
| Structure solution method | FOURIER SYNTHESIS |
| Starting model (for MR) | 1MNN.pdb without water molecules and DNA at position -1 1 2 of the MSE. |
| RMSD bond length | 0.010 |
| RMSD bond angle | 1.377 |
| Data reduction software | MOSFLM |
| Data scaling software | SCALA |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 23.570 | 1.760 |
| High resolution limit [Å] | 1.670 | 1.670 |
| Rmerge | 0.043 | 0.176 |
| Number of reflections | 51821 | |
| <I/σ(I)> | 10.5 | 3.8 |
| Completeness [%] | 99.0 | 98.3 |
| Redundancy | 4.7 | 2.7 |
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 1 mM DTT. 10mg/ml protein 1:1 molar ration with duplex DNA , 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 |
