1M9O
NMR structure of the first Zinc Binding domain of Nup475/TTP/TIS11
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D NOESY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 2 | 2D TOCSY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 3 | DQF-COSY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 4 | 3D_13C-separated_NOESY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 5 | 3D_15N-separated_NOESY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 6 | HNCACB | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 7 | 15N-HSQC-TOCSY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 8 | 13C-HCCH-TOCSY | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| 9 | HSQCJ_WEX | 1.7mM Nup475 with 2.2 Equivalent ZnCl2, 50mM d-Tris pH 5 | 90% H20, 10% D20 and 100% D20 | no salt added | 5.8 | ambient | 293 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | UNITYPLUS | 500 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| CNS, dynamical annealing protocal with molecular dynamic scheme of torsion,torsion,cartesian. Initially only distance restraints from the three Cys to the zinc were restrained. Eventually the His was also restrained and angle restraints added to make the site tetrahedral. | 442 NOE restraints, 15 Torsional angle restraints. 15 zinc site restraints, 64 Carbon chemical shifts, and 174 proton chemical restraints. | Felix |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 30 |
| Conformers Submitted Total Number | 23 |
| Representative Model | 1 (closest to the average) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | Felix | 98 | Biosym/Molecular Simulation |
| 2 | structure solution | CNS | 1.0 | Brunger |
| 3 | collection | VNMR | 6.1b | Varian |
| 4 | refinement | CNS | 1.0 | Brunger |
