1PUN

Solution Structure of the MutT Pyrophosphohydrolase Complexed with Mg(2+) and 8-oxo-dGMP, a Tightly-bound Product


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
13D_15N-separated_NOESY1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
22D NOESY1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
32D TOCSY1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
43D_13C-separated_NOESY1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
512C/13C-filtered-NOESY-HSQC1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
615N-1H IPAP-HSQC1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O, 10% D2O. 3% (w/v) bicelles (C8E5/octanol ratio 0.87)21 mM NaCl7.5ambient295
7HNCA1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
8NHCACB1MM MUTT, 1.3MM 8-OXO-DGMP, 15MM MGCL2, 4MM D11-TRIS-HCL, PH 7.5, 21MM NACL, 0.34MM NAN3.90% H2O/10% D2O21 mM NaCl7.5ambient295
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1VarianUNITYPLUS600
NMR Refinement
MethodDetailsSoftware
simulated annealing torsion angle dynamicsthe structures are based on 1746 NOE-derived distance constraints, 186 dihedral angle restraints derived from TALOS, 82 distance restraints from hydrogen bonds and 53 N-H residual dipolar couplings.NMRPipe
NMR Ensemble Information
Conformer Selection Criteriastructures with the least restraint violations
Conformers Calculated Total Number50
Conformers Submitted Total Number20
Representative Model1 (lowest energy)
Additional NMR Experimental Information
DetailsSample condition 2 was used to measure residual dipolar couplings.
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1processingNMRPipe2.1F. Delaglio
2structure solutionCNS1.1A. Brunger et al.
3data analysisTALOS1999.019.15.47Cornilecu, Delaglio and Bax
4refinementCNS1.1A. Brunger et al.