2HHI
The solution structure of antigen MPT64 from Mycobacterium tuberculosis defines a novel class of beta-grasp proteins
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 5N-edited NOESY | 2.0 mM recombinant MPT64 (15N-enriched), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 2 | NHSQC | 2.0 mM recombinant MPT64 (15N-enriched), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 3 | 13C-edited NOESY (aliphatic and aromatic) | 2.0 mM recombinant MPT64 (13C-enriched), saturated d13-MES pH 6.9 with 99% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 4 | HNCA | 2.0 mM recombinant MPT64 (15N,13C), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 5 | CBCA(CO)NH | 2.0 mM recombinant MPT64 (15N,13C), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 6 | HNCACB | 2.0 mM recombinant MPT64 (15N,13C), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 7 | HNCO | 2.0 mM recombinant MPT64 (15N,13C), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| 8 | (HCA)CO(CA)NH (Varian BioPack Software Package) | 2.0 mM recombinant MPT64 (15N,13C), saturated d13-MES pH 6.9 with 5% D2O, 0.2 mM DSS, 1mM NaN3 | no salt | 6.9 | 303 | |||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| Torsion Angle Dynamics followed by RDC refinement using CNS 1.1 | The initial structures were calculated through TAD protocol of CNS with 3761 NOE-derived distance contraints and 292 dihedral angle restraints. The final structures were refined using JNH and JCaCO Residual Dipolar Coupling (RDC) in Cartesian space with same NOE, dihedral angle restraints and 139 JNH RDCs and 114 JCaCO RDCs from stretched gel. | CNS |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 50 |
| Conformers Submitted Total Number | 24 |
| Representative Model | 1 (closest to the average) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | CNS | 1.1 | 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 |
| 2 | processing | NMRPipe | 97.027.12.56 | F. Delaglio, S. Grzesiek, G.W. Vuister, G. Zhu, J. Pfeifer, A. Bax |
| 3 | data analysis | NMRView | 5.0.4 | B.A. Johnson, R.A. Blevins |
| 4 | data analysis | Module | 1.0 | P. Dosset, J. Hus, M. Blackledge |
