1I5H

SOLUTION STRUCTURE OF THE RNEDD4 WWIII DOMAIN-RENAL BP2 PEPTIDE COMPLEX


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
13D 15N/13C-edited NOESYSample 1:1mM rNedd4 WWIII U-15N,13C; 3mM rENaC bP2 NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; Sample 2:1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; MORE DETAILED DESCRIPTION GIVEN IN BMRB 496390% H2O/10% D2O0.016.51 atm303
23D 13C F1-filtered, F3-edited NOESY1mM rNedd4 WWIII U-15N,13C; 3mM rENaC bP2 NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;100% D2O0.016.51 atm303
33D 15N/13C-edited NOESY1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;90% H2O/10% D2O0.016.51 atm303
43D 13C F1-filtered, F3-edited NOESY1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;100% D2O0.016.51 atm303
5HNCOCA_COHACA_CROSS1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;90% H2O/10% D2O0.016.51 atm303
6HNCOCA_HNHA_CROSS1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;90% H2O/10% D2O0.016.51 atm303
7J-HNHaSample 1:1mM rNedd4 WWIII U-15N,13C; 3mM rENaC bP2 NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; Sample 2:1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; MORE DETAILED DESCRIPTION GIVEN IN BMRB 496390% H2O/10% D2O0.016.51 atm303
8J-HNHa1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF;90% H2O/10% D2O0.016.51 atm303
9HNCO-TrosySample 1:1mM rNedd4 WWIII U-15N,13C; 3mM rENaC bP2 NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; Sample 2:1mM rENaC bP2 U-15N,13C; 3mM rNedd4 WWIII NA; 10 mM Na+ phosphate, pH 6.5; 0.005 mM EDTA; 0.01 mM aprotinin; 0.005 mM leupeptin; 0.005 mM AEBSF; MORE DETAILED DESCRIPTION GIVEN IN BMRB 496390% H2O/10% D2O0.016.51 atm303
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1VarianINOVA500
2VarianINOVA600
3VarianINOVA800
NMR Refinement
MethodDetailsSoftware
torsion angle dynamicsPreliminary structures were calculated using CNS1.0. These structures were then used as input into the program ARIA1.0 for noe assignment and structure refinement. These structures were calculated using 1799 unambiguous and 214 ambiguous NOEs, 14 hydrogen bond restraints, 44 dihedral angle restraints and directly refined against 33 Jhnha coupling constants.NMRPipe
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number170
Conformers Submitted Total Number15
Representative Model1 (lowest energy)
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1processingNMRPipe1.8Delaglio, F., Grzesiek, S., Vuister, G.W., Zhu, G., Pfeifer, J. and Bax, A.
2data analysisNMRVeiw4.1.2Johnson, B.A and Blevis, B.A.
3structure solutionCNS1.0Brunger, A.T., Adams, P.D., Clore, G.M., DeLano, W.L., Gros, P., Grosse-Kunstleve, R.W., Jiang, J.-S., Kuszewski, J., Nilges, M., Pannu, N.S., Read, R.J., Rice, L.M., Simonson, T. and warren, G.L.
4refinementARIA1.0Nilges, M.