2JO9

Mouse Itch 3rd WW domain complex with the Epstein-Barr virus latent membrane protein 2A derived peptide EEPPPPYED

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D CBCA(CO)NH	1.0 mM [U-100% 13C; U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
2	3D CBCANH	1.0 mM [U-100% 13C; U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
3	2D 1H-15N HSQC	1.0 mM [U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
4	3D 1H-15N NOESY	1.0 mM [U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
5	3D 1H-15N TOCSY	1.0 mM [U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
6	3D 1H-13C NOESY	1.0 mM [U-100% 13C; U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
7	3D 1H-13C TOCSY	1.0 mM [U-100% 13C; U-100% 15N] WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
8	2D 1H-1H NOESY	1.0 mM WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285
9	2D 1H-1H TOCSY	1.0 mM WW3, 3.0 mM Ligand, 20 mM Sodium Phosphate, 100 mM NaCl, 0.02% v/v Sodium Azide	90% H2O/10% D2O	0.4	6.2	ambient	285

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600
2	Bruker	DRX	800

NMR Refinement
Method	Details	Software
simulated annealing	Aria 1.2 software used, all NOEs manually assigned	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	structure solution	ARIA		Linge, O'Donoghue and Nilges
4	refinement	ARIA		Linge, O'Donoghue and Nilges

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.