2HLW
Solution Structure of the Human Ubiquitin-conjugating Enzyme Variant Uev1a
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 3D_13C-separated_NOESY | 1mM hUev1a U-15N,13C; 50 mM NaH2PO4, 250 mM NaCl, 0.9 mM DTT, 1.7 mM DSS; 90% H2O, 10% D2O | 90% H2O/10% D2O | 50 mM NaH2PO4, 250 mM NaCl | 7.0 | ambient | 303 | |
2 | 3D_15N-separated_NOESY | 1mM hUev1a U-15N,13C; 50 mM NaH2PO4, 250 mM NaCl, 0.9 mM DTT, 1.7 mM DSS; 90% H2O, 10% D2O | 90% H2O/10% D2O | 50 mM NaH2PO4, 250 mM NaCl | 7.0 | ambient | 303 | |
3 | HNHA | 1mM hUev1a U-15N,13C; 50 mM NaH2PO4, 250 mM NaCl, 0.9 mM DTT, 1.7 mM DSS; 90% H2O, 10% D2O | 90% H2O/10% D2O | 50 mM NaH2PO4, 250 mM NaCl | 7.0 | ambient | 303 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | UNITY | 600 |
2 | Varian | UNITY | 800 |
NMR Refinement | ||
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Method | Details | Software |
simulated annealing torsion angle dynamics | 2080 NOE-derived distance restraints, 56 JHNHA-derived phi restraints, 69 TALOS-derived phi restraints, 68 TALOS-derived psi restraints, 4 dNa/daN-derived psi restraints, 36 hydrogen bond restraints | CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 200 |
Conformers Submitted Total Number | 15 |
Representative Model | 1 (minimized average structure) |
Additional NMR Experimental Information | |
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Details | The structure was determined using triple-resonance NMR spectroscopy. |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | structure solution | CNS | 1.1 | Brunger |
2 | processing | NMRPipe | unknown | Delaglio |
3 | data analysis | NMRView | 5.2.2 | Johnson |
4 | collection | VNMR | 6.1 | Varian |
5 | data analysis | Sparky | 3.110 | Goddard |
6 | refinement | Procheck | 3.5.4 | Laskowski |