Novel method of protein purification for structural research. Example of ultra high resolution structure of SPI-2 inhibitor by X-ray and NMR spectroscopy.
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-1H TOCSY | 0.5 mM GmSPI 2 | 90% H2O/10% D2O | 50 | 4.5 | ambient | 298 | |
| 2 | 2D 1H-1H NOESY | 0.5 mM GmSPI 2 | 90% H2O/10% D2O | 50 | 4.5 | ambient | 298 | |
| 3 | 2D 1H-15N HSQC | 0.5 mM GmSPI 2 | 90% H2O/10% D2O | 50 | 4.5 | ambient | 298 | |
| 4 | 2D 1H-13C HSQC aliphatic | 0.5 mM GmSPI 2 | 90% H2O/10% D2O | 50 | 4.5 | ambient | 298 | |
| 5 | 2D 1H-13C HSQC aromatic | 0.5 mM GmSPI 2 | 90% H2O/10% D2O | 50 | 4.5 | ambient | 298 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | Uniform NMR System | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| molecular dynamics | CNS | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 200 |
| Conformers Submitted Total Number | 17 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | CNS | 1.3 | BRUNGER, ADAMS, CLORE, GROS, NILGES AND READ |
