SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-1H NOESY | 0.5-1.0 mM [U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 2 | 3D [1H,15N,1H]-NOESY-HSQC | 0.5-1.0 mM [U-13C; U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 3 | 3D [1H,13C,1H]-NOESY-HSQC | 0.5-1.0 mM [U-13C; U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 4 | 2D [1H,13C]-NOESY-HSQC | 0.5-1.0 mM [U-13C; U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 5 | 4D [1H,13C]-HMQC-NOESY-[1H,13C]-HMQC | 0.5-1.0 mM [U-13C; U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 6 | 4D [1H,13C]-HMQC-NOESY-[1H,15N]-HSQC | 0.5-1.0 mM [U-13C; U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 7 | 2D 1H-15N HSQC | 0.5-1.0 mM [U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 8 | 2D 1H-15N HSQC | 0.5-1.0 mM [U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| 9 | 2D 1H-15N HSQC | 0.5-1.0 mM [U-15N] N-ClpCR | 90% H2O/10% D2O | 50 | 5.5 | ambient | 305 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 800 |
| 2 | Varian | INOVA | 600 |
| 3 | Varian | INOVA | 500 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing | 200 initial structures calculated with ARIA 1.2/CNS 1.1 using NOE, dihedral, and RDC restraints, 100 lowest-energy structures were refined in AMBER 9 using the GB solvent model, 30 lowest-energy structures were deposited | ARIA |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 30 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | chemical shift assignment | ARIA | 1.2 | Linge, O'Donoghue and Nilges |
| 2 | structure solution | ARIA | 1.2 | Linge, O'Donoghue and Nilges |
| 3 | refinement | Amber | 9 | Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollm |
| 4 | model-free analysis of relaxation data | relax | 1.2 | d'Auvergne and Gooley |
| 5 | structure solution | CNS | 1.1 | Brunger, Adams, Clore, Gros, Nilges and Read |
