1K4S
HUMAN DNA TOPOISOMERASE I IN COVALENT COMPLEX WITH A 22 BASE PAIR DNA DUPLEX
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | NSLS BEAMLINE X25 |
| Synchrotron site | NSLS |
| Beamline | X25 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Spacegroup name | P 32 |
| Unit cell lengths | 73.235, 73.235, 186.632 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 50.000 - 3.200 |
| Rwork | 0.217 |
| R-free | 0.28400 * |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1a36 |
| RMSD bond length | 0.009 |
| RMSD bond angle | 1.360 * |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | CNX |
| Refinement software | CNX |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 50.000 | 3.310 |
| High resolution limit [Å] | 3.200 | 3.200 |
| Rmerge | 0.130 * | |
| Number of reflections | 17874 * | |
| <I/σ(I)> | 2.95 | |
| Completeness [%] | 85.9 | 69.5 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 8 | 16 * | PEG 8000, Tris-HCl, Na/K Phosphate, KCl, DTT, VAPOR DIFFUSION, SITTING DROP, temperature 289K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | PEG 8000 | ||
| 2 | 1 | 1 | Tris-HCl | ||
| 3 | 1 | 1 | Na/K Phosphate | ||
| 4 | 1 | 1 | KCl | ||
| 5 | 1 | 1 | DTT |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | reservoir | PEG8000 | 10 (%(w/v)) | |
| 2 | 1 | reservoir | 100 (mM) | pH8.0 | |
| 3 | 1 | reservoir | sodium potassium phosphate | 100 (mM) | pH6.2 |
| 4 | 1 | reservoir | 100 (mM) | ||
| 5 | 1 | reservoir | dithiothreitol | 10 (mM) | |
| 6 | 1 | drop | suicide duplex substrate | 0.1 (mM) | |
| 7 | 1 | drop | topo70 | 4 (mg/ml) |
