SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_13C-separated_NOESY | NMR sample: 13C, 15N-9G8 RRM at 0.6 mM Buffer conditions: 20mM CH3COONa (pH=5.5), 25mM L-ARG, 50mM L-Glu, 5mM EDTA, 10mM DTT; 90% H2O, 10% D2O | 90% H2O/10% D2O | 5.5 | ambient | 293 | ||
| 2 | 3D_15N-separated_NOESY | NMR sample: 13C, 15N-9G8 RRM at 0.6 mM Buffer conditions: 20mM CH3COONa (pH=5.5), 25mM L-ARG, 50mM L-Glu, 5mM EDTA, 10mM DTT; 90% H2O, 10% D2O | 90% H2O/10% D2O | 5.5 | ambient | 293 | ||
| 3 | 2D TOCSY | NMR sample: 13C, 15N-9G8 RRM at 0.6 mM Buffer conditions: 20mM CH3COONa (pH=5.5), 25mM L-ARG, 50mM L-Glu, 5mM EDTA, 10mM DTT; 90% H2O, 10% D2O | 90% H2O/10% D2O | 5.5 | ambient | 293 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | DRX | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| distance geometry simulated annealing | the structures are based on a total of 1138 restraints, 951 are NOE-derived distance constraints, 119 dihedral angle restraints and 68 distance restraints from hydrogen bonds | XwinNMR |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (lowest energy) |
| Additional NMR Experimental Information | |
|---|---|
| Details | The 9G8 sequence starts at K2 and finishes at R88, corresponding to K12 and R98, respectively. All the other amino-acids are part of the T7 and histidine tags. |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | collection | XwinNMR | 3.5 | |
| 2 | processing | NMRPipe | 1.0 | F. Delaglio, S. Grzesiek, G. W. Vuister, G. Zhu, J. Pfeifer and A. Bax |
| 3 | data analysis | NMRView | 5.01 | B.A.Johnson and R.A.Blevins |
| 4 | structure solution | CYANA | 2.1 | P.Guntert, C.Mumenthaler and K. Wuthrich, |
| 5 | refinement | Amber | 7.0 | Case DA, Cheatham TE 3rd, Darden T, Gohlke H, Luo R, Merz KM Jr, Onufriev A, Simmerling C, Wang B, Woods RJ |
