1RH8
Three-dimensional structure of the calcium-free Piccolo C2A-domain
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 | 0.8mM Piccolo C2A-domain U-15N, 20mM acetate (pH 6.0), 150mM NaCl, 1mM EDTA, 95% H2O, 5% D2O | 95% H2O/5% D2O | 150mM NaCl | 6.0 | ambient | 303 | |
2 | 3D_15N-separated_TOCSY | 0.8mM Piccolo C2A-domain U-15N, 20mM acetate (pH 6.0), 150mM NaCl, 1mM EDTA, 95% H2O, 5% D2O | 95% H2O/5% D2O | 150mM NaCl | 6.0 | ambient | 303 | |
3 | HNCACB | 0.8mM Piccolo C2A-domain U-15N,13C, 20mM acetate (pH 6.0), 150mM NaCl, 1mM EDTA, 95% H2O, 5% D2O | 95% H2O/5% D2O | 150mM NaCl | 6.0 | ambient | 303 | |
4 | CBCACONH | 0.8mM Piccolo C2A-domain U-15N,13C, 20mM acetate (pH 6.0), 150mM NaCl, 1mM EDTA, 95% H2O, 5% D2O | 95% H2O/5% D2O | 150mM NaCl | 6.0 | ambient | 303 | |
5 | 2D TOCSY | 0.8mM Piccolo C2A-domain U-15N, 20 mM acetate (pH 6.0), 150 mM NaCl, 1 mM EDTA | 100% D2O | 150mM NaCl | 6.0 | ambient | 303 | |
6 | 2D NOESY | 0.8mM Piccolo C2A-domain U-15N, 20 mM acetate (pH 6.0), 150 mM NaCl, 1 mM EDTA | 100% D2O | 150mM NaCl | 6.0 | ambient | 303 |
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 | The final structures were calculated with a total of 2316 experimental restraints, including 1994 NOE distance restraints, 116 H-bond restraints and 206 torsion angle restraints. | VNMR |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 1000 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (lowest energy) |
Additional NMR Experimental Information | |
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Details | This structure was determined using additional triple resonance experiments. |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | collection | VNMR | 6.1C | Varian Associates |
2 | processing | NMRPipe | 2.2 | Delaglio |
3 | data analysis | NMRView | 4.1.2 | Johson |
4 | structure solution | CNS | 0.9 | Brunger |
5 | refinement | CNS | 0.9 | Brunger |