1Y2Y

Structural Characterization of Nop10p using Nuclear Magnetic Resonance Spectroscopy

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 90% H2O, 10% D2O	90% H2O/10% D2O	110 mM	6.5	ambient	288
2	3D_13C-separated_NOESY	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 100% D2O	100% D2O	110 mM	6.5	ambient	288
3	2D TOCSY	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 100% D2O	100% D2O	110 mM	6.5	ambient	288
4	3D HCCH-TOCSY	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 100% D2O	100% D2O	110 mM	6.5	ambient	288
5	CBCANH	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 90% H2O, 10% D2O	90% H2O/10% D2O	110 mM	6.5	ambient	288
6	CBCA(CO)NH	1.5 mM 15N/13C Nop1p0p; 10 mM potassium phosphate buffer, pH 6.5; 100 mM potassium chloride; 90% H2O, 10% D2O	90% H2O/10% D2O	110 mM	6.5	ambient	288

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	500
2	Bruker	DRX	600

NMR Refinement
Method	Details	Software
refinement with simulated annealing	The structures are based on 354 intraresidue NOEs, 191 sequential NOES and 10 hydrogen bond restraints.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	30
Conformers Submitted Total Number	10
Representative Model	10 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	3.5	Bruker
2	processing	XwinNMR	3.5	Bruker
3	data analysis	AURELIA	3.0	Bruker
4	structure solution	X-PLOR	3.1	NIH
5	refinement	X-PLOR	3.1	NIH

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.