2G3Q
Solution Structure of Ede1 UBA-ubiquitin complex
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 3D_15N-separated_NOESY | 1mM 15N,13C-labeled Ede1 UBA; 1mM Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3, 90% H2O, 10% D2O | 90% H2O/10% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 | |
2 | 3D_13C-separated_NOESY | 1mM 15N,13C-labeled Ede1 UBA; 1mM Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3 | 100% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 | |
3 | 3D_15N-separated_NOESY | 1mM Ede1 UBA; 1mM 15N,13C-labeled Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3, 90% H2O, 10% D2O | 90% H2O/10% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 | |
4 | 3D_13C-separated_NOESY | 1mM Ede1 UBA; 1mM 15N,13C-labeled Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3 | 100% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 | |
5 | 3D_13C-separated_12C-edited_NOESY | 1mM 15N,13C-labeled Ede1 UBA; 1mM Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3 | 100% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 | |
6 | 3D_13C-separated_12C-edited_NOESY | 1mM Ede1 UBA; 1mM 15N,13C-labeled Ubiquitin; 20mM phosphate buffer (pH 6.0), 2mM DTT, 0.2% NaN3 | 100% D2O | 20mM Sodium Phosphate | 6.0 | 1 atm | 298 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | INOVA | 600 |
NMR Refinement | ||
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Method | Details | Software |
Torsion angle dynamics and simulated annealing | THE STRUCTURES ARE BASED ON A TOTAL OF 2489 DISTANCE RESTRAINTS, INCLUDING 2489 NOE-DERIVED DISTANCE RESTRAINTS [2069 UNAMBIGUOUS AND 420 AMBIGUOUS RESTRAINTS], 74 HYDROGEN BONDING DISTANCE RESTRAINTS, AND 161 TORSION ANGLE RESTRAINTS. | CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | The submitted conformers are the 20 structures with the lowest restraint energies, restraint violations, and RMS deviations from ideal covalent geometry |
Conformers Calculated Total Number | 80 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (closest to the average) |
Additional NMR Experimental Information | |
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Details | Additional experiments: 2D Double half filtered NOESY was performed for both D2O and H2O samples |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | refinement | CNS | 1.1 | Brunger et al. |
2 | refinement | ARIA | 1.2 | Linge and Nilges |
3 | data analysis | Felix | 98.0 | Accelrys |