2RPI
The NMR structure of the submillisecond folding intermediate of the Thermus thermophilus ribonuclease H
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 2 | 2D 1H-1H NOESY | 1.0-1.2mM protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 3 | 3D CBCA(CO)NH | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 4 | 3D HNCACB | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 5 | 3D HNCO | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 6 | 3D HNCA | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 7 | 3D HN(CO)CA | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 8 | 3D H(CCO)NH | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 9 | 3D HNHA | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 10 | 3D HCCH-TOCSY | 1.0-1.2mM [U-100% 13C] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 11 | 3D HBHA(CO)NH | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 12 | 3D 1H-15N NOESY | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 13 | 3D 1H-13C NOESY | 1.0-1.2mM [U-100% 13C] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 14 | 3D HN(CA)NNH | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| 15 | 3D HN(CA)CO | 1.0-1.2mM [U-100% 13C; U-100% 15N] protein | 100% D2O | 0.1 | 5.2 | ambient | 295 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | DRX | 500 |
| 2 | Bruker | DRX | 700 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| DGSA-distance geometry simulated annealing, torsion angle dynamics | NMRPipe | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 50 |
| Conformers Submitted Total Number | 10 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | NMRPipe | Delaglio, Zhengrong and Bax | |
| 2 | data analysis | NMRPipe | Delaglio, Zhengrong and Bax | |
| 3 | chemical shift assignment | NMRView | Johnson, One Moon Scientific | |
| 4 | peak picking | NMRView | Johnson, One Moon Scientific | |
| 5 | data analysis | TALOS | Delaglio, Zhengrong and Bax | |
| 6 | structure solution | X-PLOR NIH | Schwieters, Kuszewski, Tjandra and Clore | |
| 7 | refinement | X-PLOR NIH | Schwieters, Kuszewski, Tjandra and Clore | |
| 8 | data analysis | ProcheckNMR | Laskowski and MacArthur | |
| 9 | structure solution | MOLMOL | Koradi, Billeter and Wuthrich | |
