THREE-DIMENSIONAL STRUCTURE OF THE SYNAPTOTAGMIN 1 C2B-DOMAIN: SYNAPTOTAGMIN 1 AS A PHOSPHOLIPID BINDING MACHINE
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_15N-separated_NOESY | 0.0013M 15N, 13C C2B-Domain | 50mM Mes 2mM DTT 20mM CaCl2 5%D2O | 150mM NaCl | 6.3 | 1 atm | 303 | |
| 2 | HNHA | 0.0013M 15N, 13C C2B-Domain | 50mM Mes 2mM DTT 20mM CaCl2 5%D2O | 150mM NaCl | 6.3 | 1 atm | 303 | |
| 3 | 3D-13_15N-SEPARATED_NOESY | 1.3mM 15N, C2B-Domain | 50mM Mes 2mM DTT 20mM CaCl2 5%D2O | 150mM NaCl | 6.3 | 1 atm | 303 | |
| 4 | 1H/13C-HSQC | 1mM 10% 13C, C2B-Domain | 50mM Mes 2mM DTT 20mM CaCl2 5%D2O | 150mM NaCl | 6.3 | 1 atm | 303 | |
| 5 | 3D_15N-separated_NOESY | 0.4mM 15N, C2B-Domain | 50mM Mes 2mM DTT 0.5mM EDTA 5%D2O | 150mM NaCl | 6.3 | 1 atm | 303 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 500 |
| 2 | Varian | INOVA | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| torsion angle dynamics | the structures are based on a total of 2888 unique, conformationally significant restraints, 2492 are NOE-derived distance constraints, 214 dihedral angle restraints, 170 distance restraints from hydrogen bonds. | NMRPipe |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | STRUCTURES WITH THE MINIMUM NOE ENERGY |
| Conformers Calculated Total Number | 500 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (structure with the minimum noe energy) |
| Additional NMR Experimental Information | |
|---|---|
| Details | STRUCTURE WITH THE MINIMUM NOE ENERGY |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | NMRPipe | 1.8 | DeLaglio et al. |
| 2 | collection | VNMR | 6.1 | Varian |
| 3 | data analysis | NMRView | 4.12 | Johnson, Blevins |
| 4 | refinement | CNS | 0.9 | Brunger et al. |
