3Q23
X-ray crystal structure of the N4 mini-VRNAP and P2_7a promoter transcription initiation complex with GMPCPP and Manganese: sustrate complex II
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 |
| Collection date | 2006-11-10 |
| Detector | ADSC QUANTUM 315 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 82.834, 111.861, 276.093 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 50.000 - 1.800 |
| R-factor | 0.201 |
| Rwork | 0.200 |
| R-free | 0.23100 |
| Structure solution method | MOLECULAR REPLACEMENT |
| RMSD bond length | 0.007 |
| RMSD bond angle | 1.066 |
| Data reduction software | HKL-2000 |
| Data scaling software | HKL-2000 |
| Phasing software | CNS |
| Refinement software | REFMAC (5.2.0019) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 50.000 | 1.860 |
| High resolution limit [Å] | 1.800 | 1.800 |
| Rmerge | 0.064 | 0.545 |
| Number of reflections | 234419 | |
| <I/σ(I)> | 10.5 | |
| Completeness [%] | 99.0 | 91.5 |
| Redundancy | 6 | 4.4 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 8 | 294 | 0.1 M TRIS-HCL, 0.21 M DIBASIC AMMONIUM CITRATE, 0.09 M AMMONIUM DIHYDROGEN MONOPHOSPHATE, 25 % (DROP) 20% (RESERVOIR) PEG 3350, pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 294K |
Crystallization Reagents
| ID | crystal ID | solution ID | reagent name | concentration | details |
| 1 | 1 | 1 | TRIS-HCL | ||
| 2 | 1 | 1 | DIBASIC AMMONIUM CITRATE | ||
| 3 | 1 | 1 | AMMONIUM DIHYDROGEN MONOPHOSPHATE | ||
| 4 | 1 | 1 | PEG 3350 | ||
| 5 | 1 | 2 | TRIS-HCL | ||
| 6 | 1 | 2 | DIBASIC AMMONIUM CITRATE | ||
| 7 | 1 | 2 | AMMONIUM DIHYDROGEN MONOPHOSPHATE | ||
| 8 | 1 | 2 | PEG 3350 |
