2B9Z | pdb_00002b9z

Solution structure of FHV B2, a viral suppressor of RNAi


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
13D_15N-separated_NOESY0.2 - 1 mM 15N, 13C/15N or 2H/13C/15N labelled B2 protein in 50 mM sodium phosphate pH 6.3, 50 mM sodium chloride, 1 mM DTT, 90% H2O / 10% D2O90% H2O/10% D2O50 mM NaPi, 50 mM NaCl6.31 atm295
23D_13C-separated_NOESY0.2 - 1 mM 13C/15N labelled B2 protein in 50 mM sodium phosphate pH 6.3, 50 mM sodium chloride, 1 mM DTT100% D2O50 mM NaPi, 50 mM NaCl6.31 atm295
3HNHA0.2 - 1 mM 15N, 13C/15N or 2H/13C/15N labelled B2 protein in 50 mM sodium phosphate pH 6.3, 50 mM sodium chloride, 1 mM DTT, 90% H2O / 10% D2O90% H2O/10% D2O50 mM NaPi, 50 mM NaCl6.31 atm295
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerDRX500
2BrukerDRX600
3BrukerDRX900
NMR Refinement
MethodDetailsSoftware
molecular dynamics, simulated annealingThe experimentally determined distance, dihedral and dipolar coupling restraints were applied in a simulated annealing protocol using ARIA and CNS. Non-crystallographic symmetry (NCS) restraints were used to enforce the dimer symmetry. The final ensemble of NMR structures was refined in a shell of water molecules.XwinNMR
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number100
Conformers Submitted Total Number10
Representative Model1 (lowest energy)
Additional NMR Experimental Information
DetailsDistance restraints were derived from 15N- or 13C-resolved 3D NOESY. HN-N residual dipolar couplings were measured using a spin-state-selective 1H,15N correlation experiment in dilute liquid crystalline medium. Restraints for the backbone angles phi and psi were derived from TALOS. Stereospecific assignments of Leu, Val methyl groups were obtained using a 10% fractionally 13C-labelled sample.
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1collectionXwinNMR3.5
2processingNMRPipeDelaglio et al.
3data analysisNMRView5Johnson et al.
4data analysisTALOSCornilescu et al.
5structure solutionARIA1.2Linge et al.
6structure solutionCNS1.1Brunger et al.
7refinementCNS1.1Brunger et al.