2POA
Schistosoma mansoni Sm14 Fatty Acid-Binding Protein: improvement of protein stability by substitution of the single Cys62 residue
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | HNCA | U-15N, 13C; 50 mM phosphate buffer pH 6.3; 0.05% sodium Azide; 95% H2O, 5% D2O. | 95% H2O/5% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 | |
2 | HNCOCA | U-15N, 13C; 50 mM phosphate buffer pH 6.3; 0.05% sodium Azide; 95% H2O, 5% D2O. | 95% H2O/5% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 | |
3 | HNCACB | U-15N, 13C; 50 mM phosphate buffer pH 6.3; 0.05% sodium Azide; 95% H2O, 5% D2O. | 95% H2O/5% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 | |
4 | CBCACONH | U-15N, 13C; 50 mM phosphate buffer pH 6.3; 0.05% sodium Azide; 95% H2O, 5% D2O. | 95% H2O/5% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 | |
5 | 3D_13C-separated_NOESY | U-15N, 13C; 100% D2O | 100% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 | |
6 | 3D_15N-separated_NOESY | U-15N, 13C; 50 mM phosphate buffer pH 6.3; 0.05% sodium Azide; 95% H2O, 5% D2O. | 95% H2O/5% D2O | 50 mM phosphate buffer | 6.3 | ambient | 293 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | INOVA | 500 |
2 | Varian | INOVA | 600 |
3 | Bruker | AVANCE | 600 |
NMR Refinement | ||
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Method | Details | Software |
torsion angle dynamics using dyana software and further minimization with discover module of the insight package | minimization were conducted with 5000 iterations using steepest descents algorithim until the maximum derivative is less than 1.000 kcal/A followed by 50000 iterations using conjugate gradients until the maximum derivative is less than 0.00100cal/A | DYANA |
NMR Ensemble Information | |
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Conformer Selection Criteria | target function |
Conformers Calculated Total Number | 40 |
Conformers Submitted Total Number | 17 |
Representative Model | 17 (fewest violations) |
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 | DYANA | 1.5 | Guntert, Braun and Wuthrich |
2 | refinement | Discover | ||
3 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
4 | data analysis | NMRView | Johnson, One Moon Scientific |