SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D 1H-15N NOESY | 1.5 mM [U-13C; U-15N] protein, 150 mM sodium chloride, 10 mM DTT, 0.05 % sodium azide, 0.2 mM PMSF, 1 mM EDTA | 93% H2O/7% D2O | 7.0 | ambient | 298 | ||
| 2 | 3D 1H-13C NOESY aliphatic | 1.5 mM [U-13C; U-15N] protein, 150 mM sodium chloride, 10 mM DTT, 0.05 % sodium azide, 0.2 mM PMSF, 1 mM EDTA | 93% H2O/7% D2O | 7.0 | ambient | 298 | ||
| 3 | 2D 1H-13C HSQC aromatic | 1.5 mM [U-13C; U-15N] protein, 150 mM sodium chloride, 10 mM DTT, 0.05 % sodium azide, 0.2 mM PMSF, 1 mM EDTA | 93% H2O/7% D2O | 7.0 | ambient | 298 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 1000 |
| 2 | Bruker | AVANCE | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing | X-PLOR NIH | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | structure solution | X-PLOR NIH | Schwieters, Kuszewski, Tjandra and Clore | |
| 2 | refinement | Amber | Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollman | |
| 3 | chemical shift assignment | CARA | Keller and Wuthrich | |
| 4 | structure solution | CYANA | 2.1 | Guntert, Mumenthaler and Wuthrich |
| 5 | structure solution | UNIO'10 | 2.02 | (UNIO)Torsten Herrmann |
| 6 | processing | TopSpin | Bruker Biospin | |
