1XSC
Structure of the nudix enzyme AP4A hydrolase from homo sapiens (E63A mutant) in complex with ATP
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_15N-separated_NOESY | 1.0mM human AP4A hydrolase, 13C 15N ATP | 20mM MgCl2, 20mM imidazole, pH 6.5, 90% H2O 10% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP (13C,15N labelled) | 51mM | 6.5 | ambient | 293 | |
| 2 | 13C noesy over Aromatic/Anomeric | 1.0mM human AP4A hydrolase, 13C 15N ATP | 20mM MgCl2, 20mM imidazole, pH 6.5, 100% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP (13C, 15N labelled) | 51mM | 6.5 | ambient | 293 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 600 |
| 2 | Varian | INOVA | 500 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| CNS from a starting structure with ATP 20A from binding site, Refine against NOE/CACB no RAMA potential | CNS | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | target function |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 25 |
| Representative Model | 1 (lowest energy) |
| Additional NMR Experimental Information | |
|---|---|
| Details | This calcualtion includes ATP constraints. Slow cooling, Lower temperature (20000K rather than 50000K) ATP starts 20A from lowest energy conformer Residues 1 and 2 are changed from GP to Alanine in the calculations. These are from the non native precission cleavage site and are unstructrued. |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | CNS | 1.0 | |
