2L15
Solution Structure of Cold Shock Protein CspA Using Combined NMR and CS-Rosetta method
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 2 | 3D HNCO | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 3 | 3D HNcaCO | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 4 | 3D HNCA | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 5 | 3D HN(CO)CA | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 6 | 3D HNCACB | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| 7 | 3D HN(COCA)CB | 0.2 mM [U-100% 2H; 100% 13C; U-100% 15N] ILV Methyl 1H CspA-1 | 95% H2O/5% D2O | 50 | 6.0 | ambient | 20 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing, molecular dynamics | AutoAssign | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the best score calculted by CS-Rosetta |
| Conformers Calculated Total Number | 10000 |
| Conformers Submitted Total Number | 10 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | chemical shift assignment | AutoAssign | Zimmerman, Moseley, Kulikowski and Montelione | |
| 2 | data analysis | Sparky | Goddard and Kneller | |
| 3 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 4 | data analysis | PSVS | Bhattacharya, Tejero and Montelione | |
| 5 | structure solution | CS-ROSETTA | Bax and Yang | |
| 6 | refinement | CS-ROSETTA | Bax and Yang | |
