1WR1

The complex structure of Dsk2p UBA with ubiquitin

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	4D_13C/15N-separated_NOESY	U-15N, 13C ubiquitin + DSK2-UBA complex (0.9mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
2	3D_15N-separated_NOESY	U-15N, 13C ubiquitin + DSK2-UBA complex (0.9mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
3	3D_13C-separated_NOESY	U-15N, 13C ubiquitin + DSK2-UBA complex (0.9mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
4	3D_13C-edited/13C-filtered-noesy	U-15N, 13C ubiquitin + DSK2-UBA complex (0.9mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
5	4D_13C/15N-separated_NOESY	U-15N, 13C DSK2-UBA + ubiquitin complex (1.0mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
6	3D_15N-separated_NOESY	U-15N, 13C DSK2-UBA + ubiquitin complex (1.0mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
7	3D_13C-separated_ROESY	U-15N, 13C DSK2-UBA + ubiquitin complex (1.0mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298
8	3D_13C-edited/13C-filtered-noesy	U-15N, 13C DSK2-UBA + ubiquitin complex (1.0mM)	20mM Phosphate buffer (pH 6.8); 5mM potassium chloride; 1mM EDTA; 5% D2O	20mM potassium phosphate, 5mM potassium chloride	6.8		298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	800
2	Varian	INOVA	900

NMR Refinement
Method	Details	Software
torsion angle dynamics, simulated annealing, molecular dynamics, energy minimization		XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	16 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	3.5
2	collection	VNMR	6.1C
3	processing	NMRPipe	2.1	Delaglio
4	data analysis	Sparky	3.1.0.0	Goddard
5	structure solution	CYANA	2.0.17	Guntert
6	refinement	Amber	7	Pearlman

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.