THE NMR SOLUTION STRUCTURE OF THE COMPLEX FORMED BETWEEN ALPHA-BUNGAROTOXIN AND AN 18MER COGNATE PEPTIDE
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_15N-separated_TOCSY | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| 2 | 3D HNHA | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| 3 | 3D_15N-separated_NOESY | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| 4 | 2D_TOCSY | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| 5 | 2D_NOESY | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| 6 | 2D 1H-15N HSQC | 2.1 mM alpha-Bungarotoxin/alpha18mer complex, alpha18mer is 15N labeled, 50 mM perdeuterated potassium acetate buffer (pH 4.0) with 5% D2O and 0.05% sodium azide | 95% H2O/5% D2O | 4.0 | ambient | 308 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| distance geometry and simulated annealing | XwinNMR | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | The submitted conformer models are the 20 structures with the lowest energy from 120 acceptable structures which were from 300 total calculated structures and had no restraint violations. The best representative conformer is the lowest potential energy structure of the ensemble. |
| Conformers Calculated Total Number | 300 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 8 (it is the lowest potential energy structure of the ensemble) |
| Additional NMR Experimental Information | |
|---|---|
| Details | The authors recommend using the Graphics program, MOLMOL, to view this ensemble of 20 structures. |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | collection | XwinNMR | 2.1 | Bruker |
| 2 | processing | NMRPipe | 1.8 | Delaglio, F. et al. |
| 3 | data analysis | Sparky | 3.95 | Goddard, T.D. and Kneller, D.G. |
| 4 | refinement | CNSsolve | 1.0 | Brunger, A.T. et al. |
