2KDV
Solution structure of RNA Pyrophosphohydrolase RppH from Escherichia coli
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 50mM sodium acetate-1, 20mM sodium chloride-2, 40mM DTT-3, 10mM arginine-4, 10mM glutamic-5, 0.8mM [U-13C; U-15N] RppH protein-6 | 90% H2O/10% D2O | 90 | 5 | ambient | 303 | |
| 2 | 3D 1H-15N NOESY | 50mM sodium acetate-1, 20mM sodium chloride-2, 40mM DTT-3, 10mM arginine-4, 10mM glutamic-5, 0.8mM [U-13C; U-15N] RppH protein-6 | 90% H2O/10% D2O | 90 | 5 | ambient | 303 | |
| 3 | 3D 1H-13C NOESY | 50mM sodium acetate-1, 20mM sodium chloride-2, 40mM DTT-3, 10mM arginine-4, 10mM glutamic-5, 0.8mM [U-13C; U-15N] RppH protein-6 | 90% H2O/10% D2O | 90 | 5 | ambient | 303 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 600 |
| 2 | Bruker | AVANCE | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing | NMRPipe | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 21 |
| Representative Model | 1 (minimized average) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 2 | data analysis | NMRView | Johnson, One Moon Scientific | |
| 3 | collection | XwinNMR | Bruker Biospin | |
| 4 | refinement | Amber | Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollm | |
| 5 | structure solution | CYANA | Guntert, Mumenthaler and Wuthrich | |
