2NLN
Solution Structure of Calcium-free Rat Beta-parvalbumin
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 | 4 mM oncomodulin, U-15N, 0.15 M NaCl, 0.01 M Mes, pH 6.0, 90% H2O, 10% D2O | 90% H2O/10% D2O | 0.15 M NaCl, 0.01 M Mes | 6.0 | 1 atm | 293 | |
2 | 3D_13C-separated_NOESY | 4 mM oncomodulin, U-15N, 13C, 0.15 M NaCl, 0.01 M Mes, pH 6.0, 90% H2O, 10% D2O | 90% H2O/10% D2O | 0.15 M NaCl, 0.01 M Mes | 6.0 | 1 atm | 293 | |
3 | 15N-IPAP-HSQC | 4 mM oncomodulin, U-15N, 0.15 M NaCl, 0.01 M Mes, pH 6.0, 13 mg/mL Pf1 bacteriophage, 90% H2O, 10% D2O | 90% H2O/10% D2O | 0.15 M NaCl, 0.01 M Mes | 6.0 | 1 atm | 293 | |
4 | 15N-HSQC | 4 mM oncomodulin, U-15N, 0.15 M NaCl, 0.01 M Mes, pD 6.2 | 100% D2O | 0.15 M NaCl, 0.01 M Mes | 6.0 | 1 atm | 293 | |
5 | HNHA | 4 mM oncomodulin, U-15N, 0.15 M NaCl, 0.01 M Mes, pH 6.0, 90% H2O, 10% D2O | 90% H2O/10% D2O | 0.15 M NaCl, 0.01 M Mes | 6.0 | 1 atm | 293 |
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 |
simulated annealing using torsion angle molecular dynamics | The structures are based on a total of 2961 restraints: 2553 are NOE-derived distance constraints, 241 are dihedral angle restraints, 74 are distance restraints derived from putative hydrogen bonds, and 93 are angle restraints based on residual dipolar couplings | VNMR |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 50 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (closest to the average) |
Additional NMR Experimental Information | |
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Details | This structure was determined using: 1. distance restraints collected from 3D NOESY experiments, 2. dihedral angle restraints based on coupling constants (HNHA), and chemical shift data (CSI, TALOS), 3. hydrogen-bond restraints (collected from H-D exchange measurements), 4. 1H-15N residual dipolar couplings |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | collection | VNMR | varian | |
2 | processing | NMRPipe | Delaglio, F. et al. | |
3 | data analysis | Sparky | Goddard T.D., Kneller, D.G. | |
4 | structure solution | ARIA | 2.0a | Nilges, M. et al. |
5 | structure solution | CNS | 1.1 | Brunger, A.T. et al. |
6 | refinement | CNS | 1.1 | Brunger, A.T. et al. |