1EJ5
SOLUTION STRUCTURE OF THE AUTOINHIBITED CONFORMATION OF WASP
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 | 1.6 mM protein U-15N,13C; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 | |
2 | 3D_15N-separated_NOESY | 1.6 mM protein U-15N,13C; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 | |
3 | 4D_13C-separated_NOESY | 1.6 mM protein U-15N,13C; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 | |
4 | 4D_13C/15N-separated_NOESY | 1.6 mM protein U-15N,13C; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 | |
5 | HNHA | 1.6 mM protein U-15N; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 | |
6 | HCCH-TOCSY | 1.6 mM protein U-15N,13C; 25 mM phosphate buffer pH 6.5, 50 mM NaCl, 1 mM EDTA, 2 mM DTT | 90% H2O/10% D2O | 75mM | 6.5 | ambient | 298 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | INOVA | 600 |
2 | Varian | INOVA | 500 |
NMR Refinement | ||
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Method | Details | Software |
simulated annealing, torsion angle dynamics | The structures are based on a total of 2713 restraints, 2494 are NOE-derived distance constraints, 182 dihedral angle restraints, 31 distance restraints from hydrogen bonds. | CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the least restraint violations |
Conformers Calculated Total Number | 100 |
Conformers Submitted Total Number | 20 |
Representative Model | 14 (lowest energy) |
Additional NMR Experimental Information | |
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Details | The structure was determined using standard 3D and 4D heteronuclear techniques. |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | refinement | CNS | 0.3 | Brunger, Nilges |
2 | refinement | XPLOR | 3.851 | Brunger |
3 | processing | NMRPipe | 1.7 | Delaglio |
4 | data analysis | NMRView | 2.1.2 | Jonhson, Blevins |
5 | collection | VNMR | 6.1b | VARIAN |
6 | refinement | ARIA | 1 | Nilges |