SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_15N-separated_NOESY | 0.4mM U-15N | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 95% H2O, 5% D2O | 7.0 | ambient | 295 | ||
| 2 | 3D_13C-separated_NOESY | 0.6mM U-15N, 13C | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 95% H2O, 5% D2O | 7.0 | ambient | 295 | ||
| 3 | HNHA | 0.4mM U-15N | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 95% H2O, 5% D2O | 7.0 | ambient | 295 | ||
| 4 | 2D NOESY | 0.4mM U-15N | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 100% D2O | 7.0 | ambient | 295 | ||
| 5 | 2D NOESY | 0.4mM U-15N | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 95% H2O, 5% D2O | 7.0 | ambient | 295 | ||
| 6 | 1H-15N HSQC | 0.4mM U-15N | 10mM sodium phosphate, 50mM sodium chloride, 2mM EDTA, 0.02%(w/w) sodium azide, 100% D2O | 7.0 | ambient | 295 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 500 |
| 2 | Bruker | DRX | 600 |
| 3 | Bruker | AVANCE | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| torsion angle dynamics, simulated annealing | The structures are based on a total of 2074, 1738 are non-redundant NOE-derived distance constraints, 260 dihedral angle restraints, 76 distantce restraints from hydrogen bonds. | XwinNMR |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with acceptable covalent geometry |
| Conformers Calculated Total Number | 194 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (closest to the average) |
| Additional NMR Experimental Information | |
|---|---|
| Details | The structure was determined using triple-resonance NMR spectroscopy. Constrants for backbone Phi and Psi were also obtained from program TALOS using backbone chemical shifts. |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | collection | XwinNMR | 3.5 | Bruker, Biospin |
| 2 | processing | XwinNMR | 3.5 | Bruker, Biospin |
| 3 | data analysis | XEASY | 1.3 | Bartels et al |
| 4 | structure solution | CYANA | 1.0.6 | Guntert et al |
| 5 | structure solution | X-PLOR | 2.9.3 | Schwieters |
| 6 | refinement | X-PLOR | 2.9.3 | Schwieters |
