2L0D
Solution NMR Structure of putative cell surface protein MA_4588 (272-376 domain) from Methanosarcina acetivorans, Northeast Structural Genomics Consortium Target MvR254A
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 2 | 2D 1H-13C HSQC | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 3 | 3D HNCO | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 4 | 3D CBCA(CO)NH | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 5 | 3D HNCACB | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 6 | 3D 1H-13C arom NOESY | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 7 | 3D 1H-13C aliph NOESY | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 8 | 3D 1H-15N NOESY | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 9 | 4D 1H-13C-13C-1H HMQC-NOESY-HMQC | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 100% D2O | 6.5 | ambient | 298 | ||
| 10 | 3D HBHA(CO)NH | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 11 | 3D HCCH-TOCSY | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 12 | 3D C(CO)NH | 1.1 mM [U-100% 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| 13 | 2D 1H-13C HSQC | 1.1 mM [5% biosynthetically-directed 13C; U-100% 15N] MvR254A, 100 mM NaCl, 20 mM MES, 5 mM CaCl2, 0.05 % NaN3 | 95% H2O/5% D2O | 6.5 | ambient | 298 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 750 |
| 2 | Varian | INOVA | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| molecular dynamics and simulated annealing | CNS | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy |
| Conformers Calculated Total Number | 40 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | CNS | Brunger, Adams, Clore, Gros, Nilges and Read | |
| 2 | structure solution | CNS | Brunger, Adams, Clore, Gros, Nilges and Read | |
| 3 | geometry optimization | CNS | Brunger, Adams, Clore, Gros, Nilges and Read | |
| 4 | refinement | CYANA | 3.0 | Guntert, Mumenthaler and Wuthrich |
| 5 | geometry optimization | CYANA | 3.0 | Guntert, Mumenthaler and Wuthrich |
| 6 | structure solution | CYANA | 3.0 | Guntert, Mumenthaler and Wuthrich |
| 7 | data analysis | AutoStructure | 2.1 | Huang, Tejero, Powers and Montelione |
| 8 | data analysis | AutoAssign | 2.1 | Zimmerman, Moseley, Kulikowski and Montelione |
| 9 | chemical shift assignment | AutoAssign | 2.1 | Zimmerman, Moseley, Kulikowski and Montelione |
| 10 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 11 | data analysis | XEASY | Bartels et al. | |
| 12 | peak picking | XEASY | Bartels et al. | |
| 13 | chemical shift assignment | XEASY | Bartels et al. | |
| 14 | collection | TopSpin | Bruker Biospin | |
| 15 | collection | VnmrJ | Varian | |
| 16 | chemical shift assignment | PINE | Bahrami, Markley, Assadi, and Eghbalnia | |
| 17 | data analysis | Sparky | Goddard | |
| 18 | geometry optimization | TALOS+ | Shen, Cornilescu, Delaglio and Bax | |
| 19 | geometry optimization | PALES | PALES (Zweckstetter, Bax) | |
| 20 | geometry optimization | REDCAT | Valafar, Prestegard | |
| 21 | refinement | AutoStructure | 2.1 | Huang, Tejero, Powers and Montelione |
| 22 | refinement | Xplor-NIH | Schwieters, Kuszewski, Tjandra and Clore | |
