2KR0

Solution structure of the proteasome ubiquitin receptor Rpn13


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
12D 1H-15N HSQC0.1 mM [U-15N] protein, 0.1 mM [U-15N; U-50% 2H] protein90% H2O/10% D2O20 mM phosphate, 50 mM NaCl6.5298
23D 1H-15N NOESY0.1 mM [U-15N] protein, 0.1 mM [U-15N; U-50% 2H] protein90% H2O/10% D2O20 mM phosphate, 50 mM NaCl6.5298
32D 1H-15N HSQC0.1 mM [U-15N] protein, 0.1 mM [U-15N; U-50% 2H] protein90% H2O/10% D2O20 mM phosphate, 50 mM NaCl6.5298
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1VarianINOVA800
2VarianINOVA900
3BrukerAVANCE700
NMR Refinement
MethodDetailsSoftware
simulated annealingThe structure of two functional domain of the protein is calculated based on their NOE-derived distance constraints, hydrogen bonds and dihedral angle restraints. The paramagnetic relaxation enhancement from spin-labeling experiments and inter-domain NOE help define the structure of full protein.VnmrJ
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number50
Conformers Submitted Total Number31
Representative Model1 (lowest energy)
Additional NMR Experimental Information
DetailsThe Spectra took in Bruker Advance 700 MHz are a set of spin-labeling experiments to achieve PRE data.
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1collectionVnmrJVarian
2collectionTopSpinBruker Biospin
3processingNMRPipe2.5Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4chemical shift assignmentXEASY6.4Bartels et al.
5data analysisXEASY6.4Bartels et al.
6peak pickingXEASY6.4Bartels et al.
7chemical shift assignmentCARA1.8.4Keller and Wuthrich
8data analysisCARA1.8.4Keller and Wuthrich
9refinementX-PLOR NIHSchwieters, Kuszewski, Tjandra and Clore
10structure solutionX-PLOR NIHSchwieters, Kuszewski, Tjandra and Clore