1ZLL

NMR Structure of Unphosphorylated Human Phospholamban Pentamer


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
1TROSY-HNCA, HNCACB, HNCACO0.2 mM Phospholamban Pentamer, U-15N, 13C, 85% 2H, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
2measurement of 1H-15N, 13C'-13CA, and 13C'-15N RDCs using 3D HNCO based experiments0.05 mM Phospholamban Pentamer, U-15N,13C, 85% 2H, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM DPC, 4% stretched polyacrylamide gel, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
3HCCONH, CCONH0.2 mM Phospholamban Pentamer, U-15N,13C, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM 2H-DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
415N-edited NOESY, 13C-edited NOESY0.2 mM Phospholamban Pentamer, U-15N,13C, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM 2H-DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
513C-HSQC for methyl stereospecific assignment0.2 mM Phospholamban Pentamer, 10% 13C-labeled, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
6Double 13C-filtered, 15N-edited NOESY0.2 mM Phospholamban Pentamer, 50% U-15N,13C and 50% unlabeled, 25mM Sodium Phosphate buffer, 50 mM BME, 200 mM 2H-DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
73D 15N-edited NOESY0.2 mM Phospholamban Pentamer, 50% U-15N, U-2H and 50% unlabeled; 25mM Sodium Phosphate buffer, 50 mM BME, 200 mM 2H-DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
82D spin-echo difference experiments for measuring 3-bond J(NCg) and J(CCg) couplings0.2 mM Phospholamban Pentamer, U-15N,13C, 25 mM Sodium Phosphate buffer, 50 mM BME, 200 mM 2H-DPC, 95% H2O, 5% D2O95% H2O/5% D2O25 mM Sodium Phosphate6.0ambient303
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerAVANCE500
2BrukerDMX600
3BrukerAVANCE750
NMR Refinement
MethodDetailsSoftware
Simulated Annealing, Molecular Dynamics1. Monomer structure was determined with simulated annealing protocol that refines all intramonomer NOEs, sidechain J-couplings, and backbone RDCs. 2. Pentamer is calculated with simulated-annealing protocol that treats individual monomer backbones as rigid bodies while satisfying all intermonomer NOEs.NMRPipe
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number30
Conformers Submitted Total Number20
Representative Model9 (fewest violations)
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1processingNMRPipe2.2Delaglio et al.
2refinementXPLOR-NIH2.9.9Schwieters et al.