1KFT
Solution Structure of the C-Terminal domain of UvrC from E-coli
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D NOESY | 1mM UvrC-CTD 15N labelled 300mM Nacl, 50mM Phosphate buffer 0.2mM complete protease inhibitor 95% H2O, 5% D2O, pH 6.8 | 95% H2O/5% D2O | 300mM Nacl, 50mM phosphate buffer | 6.8 | atmospheric atm | 300 | |
| 2 | 3D_15N-separated_NOESY | 1mM UvrC-CTD 15N labelled 300mM Nacl, 50mM Phosphate buffer 0.2mM complete protease inhibitor 95% H2O, 5% D2O, pH 6.8 | 95% H2O/5% D2O | 300mM Nacl, 50mM phosphate buffer | 6.8 | atmospheric atm | 300 | |
| 3 | 3D_13C-separated_NOESY | 1mM UvrC-CTD 15N 13C doublly labelled 300mM Nacl, 50mM Phosphate buffer 0.2mM complete protease inhibitor 95% H2O, 5% D2O, pH 6.8 | 95% H2O/5% D2O | 300mM Nacl, 50mM phosphate buffer | 6.8 | atmospheric atm | 300 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AMX | 500 |
| 2 | Bruker | DRX | 600 |
| 3 | Varian | INOVA | 750 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| A simulated annealing protocol in Cartesian space was used. Final refinement was done with explicit water. | Calculation was done on the basis of 1326 unambiguous and 60 ambiguous distance restraints with the inclusion of 68 dihedral angle constraints. | NMRPipe |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | no NOE distance violations > 0.5A and lowest energies after refining the best 50 of the initial 200 in explicit water |
| Conformers Calculated Total Number | 200 |
| Conformers Submitted Total Number | 22 |
| Representative Model | 5 (fewest violations,lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | NMRPipe | version 1.8 | Delaglio, F., Grzesiek, S., Vuister, G., Zhu, G., Pfeifer, J., and Bax, A. |
| 2 | data analysis | NMRView | 4.1.3 | Bruce A. Johnson |
| 3 | structure solution | ARIA | 1.0 | S.,Jens Linge, Michael Nilges |
| 4 | structure solution | CNS | 1.0 | A.T.Brunger, P.D.Adams, G.M.Clore, W.L.DeLano,P.Gros, R.W.Grosse-Kunstleve, J.-S.Jiang,J.Kuszewski, M.Nilges, N.S.Pannu, R.J.Read, L.M.Rice, T.Simonson, G.L.Warren. |
| 5 | refinement | CNS | 1.0 | A.T.Brunger, P.D.Adams, G.M.Clore, W.L.DeLano,P.Gros, R.W.Grosse-Kunstleve, J.-S.Jiang,J.Kuszewski, M.Nilges, N.S.Pannu, R.J.Read, L.M.Rice, T.Simonson, G.L.Warren. |
