Solution NMR structure of glutamine amido transferase subunit of gaunosine monophosphate synthetase from Methanocaldococcus jannaschii
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 2 | 2D 1H-13C HSQC | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 3 | 3D HNCA | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 4 | 3D HN(CO)CA | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 5 | 3D CBCA(CO)NH | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 6 | 3D HNCACB | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 7 | 3D 1H-15N NOESY | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 8 | 3D 1H-15N TOCSY | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 9 | 3D HNHA | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 10 | 3D HCCH-TOCSY | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| 11 | 3D HCCH-COSY | 0.7mM [U-99% 15N] GATase subunit-1, 10% D2O-2, 90% H2O-3, 1mM PMSF-4, 20mM potassium phosphate-5, 0.1mM EDTA-6, 2mM DTT-7, 0.01% sodium azide-8 | 90% H2O/10% D2O | 7.0 | ambient | 303 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 700 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| na | NMRPipe | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | target function |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (target function) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 2 | peak picking | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 3 | chemical shift assignment | CCPN | CCPN | |
| 4 | data analysis | CCPN | CCPN | |
| 5 | peak picking | CCPN | CCPN | |
| 6 | structure solution | CCPN | CCPN | |
| 7 | structure solution | Cyana-3.0 | Guntert, Mumenthaler and Wuthrich | |
| 8 | data analysis | TALOS+ | Cornilescu, Delaglio and Bax | |
| 9 | geometry optimization | TALOS+ | Cornilescu, Delaglio and Bax | |
| 10 | structure solution | TALOS+ | Cornilescu, Delaglio and Bax | |
| 11 | data analysis | Procheck | Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Thornton | |
| 12 | geometry optimization | Procheck | Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Thornton | |
| 13 | structure solution | Procheck | Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Thornton | |
| 14 | data analysis | ProcheckNMR | Laskowski and MacArthur | |
| 15 | geometry optimization | ProcheckNMR | Laskowski and MacArthur | |
| 16 | structure solution | ProcheckNMR | Laskowski and MacArthur | |
| 17 | data analysis | CSI | Canadian Institutes of Health Research (CIHR Group) | |
| 18 | structure solution | CSI | Canadian Institutes of Health Research (CIHR Group) | |
| 19 | structure solution | MOLMOL | Koradi, Billeter and Wuthrich | |
| 20 | data analysis | MOLMOL | Koradi, Billeter and Wuthrich | |
| 21 | processing | NMR-STAR | Hall, S. R. | |
| 22 | data analysis | PSVS | Bhattacharya and Montelione | |
| 23 | structure solution | PSVS | Bhattacharya and Montelione | |
| 24 | geometry optimization | PSVS | Bhattacharya and Montelione | |
| 25 | refinement | Cyana-3.0 | ||
