SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D 1H-15N NOESY | 1.75 mM [U-100% 13C; U-100% 15N] protein, 90%H2O/10% D2O | 90% H2O/10% D2O | 0.1 | 6.8 | ambient | 298 | |
| 2 | 3D 1H-13C NOESY | 1.25 mM [U-100% 13C; U-100% 15N; 80% 2H] protein | 90% H2O/10% D2O | 0.1 | 6.8 | ambient | 298 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 600 |
| 2 | Bruker | AVANCE | 600 |
| 3 | Bruker | AVANCE | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| torsion angle dynamics, simulated annealing | Following Nederveen AJ et al., Proteins 2005, 59: 662-672 but in presence of a 10angstrom explicit water shell, final minimization in presence of 4.5 Angstrom explicit water shell | NMRView |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the least restraint violations |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (fewest violations) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | chemical shift assignment | NMRView | Johnson, One Moon Scientific | |
| 2 | data analysis | NMRView | Johnson, One Moon Scientific | |
| 3 | data analysis | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 4 | chemical shift assignment | CYANA | 2.3 | Guntert, Mumenthaler and Wuthrich |
| 5 | geometry optimization | CYANA | 2.3 | Guntert, Mumenthaler and Wuthrich |
| 6 | structure solution | X-PLOR NIH | 2.17.0 | Schwieters, Kuszewski, Tjandra and Clore |
| 7 | refinement | X-PLOR NIH | 2.17.0 | Schwieters, Kuszewski, Tjandra and Clore |
